Flea protease proteins and uses thereof

ABSTRACT

The present invention relates to flea serine protease proteins, aminopeptidase proteins and flea cysteine protease proteins; to flea serine protease, aminopeptidase and cysteine protease nucleic acid molecules, including those that encode such proteins; to antibodies raised against such proteins; and to compounds that inhibit flea serine protease, aminopeptidase and/or cysteine protease activities. The present invention also includes methods to obtain such proteins, nucleic acid molecules, antibodies, and inhibitors. Also included in the present invention are therapeutic compositions comprising such proteins, nucleic acid molecules, antibodies, and/or inhibitors as well as the use of such therapeutic compositions to protect a host animal from flea infestation.

CROSS-REFERENCE TO RELATE APPLICATIONS

The present invention is a continuation-in-part of U.S. patentapplication Ser. No. 08/484,211. (issued as U.S. Pat. No. 5,972,645 onOct. 26, 1999), Ser. No. 08/482,130 (issued as U.S. Pat. No. 5,962,257on Oct. 5, 1999), Ser. Nos. 08/485,443, and 08/485,455 (issued as U.S.Pat. No. 5,712,143 on Jan. 27, 1998), each of which was filed on Jun. 7,1995 and each of which is a continuation-in-part of U.S. patentapplication Ser. No. 08/326,773, filed Oct. 18, 1994 which issued asU.S. Pat. No. 5,766,609 on Jun. 16, 1998, which is acontinuation-in-part of U.S. patent application Ser. No. 07/806,482,filed Dec. 13, 1991, which issued as U.S. Pat. No. 5,356,622 on Oct. 18,1994. The present application is also a continuation-in-part of U.S.patent application Ser. No. 08/326,773, ibid., as well as ofPCT/US95/14442, the international filing date of which is Oct. 18, 1995.PCT/US95/14442 claims priority to U.S. patent application Ser. Nos.08/326,773, 08/484,211, 08/482,130, 08/485,443 and 08/485,455, ibid.Each of the applications referred to in this section is incorporatedherein by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to novel flea protease proteins and theiruse to reduce flea infestation of animals. The present invention alsorelates to the use of anti-flea protease antibodies and other compoundsthat reduce flea protease activity to reduce flea infestation ofanimals.

BACKGROUND OF THE INVENTION

Fleas, which belong to the insect order Siphonaptera, are obligateectoparasites for a wide variety of animals, including birds andmammals. Flea infestation of animals is of health and economic concernbecause fleas are known to cause and/or transmit a variety of diseases.Fleas cause and/or carry infectious agents that cause, for example, fleaallergy dermatitis, anemia, murine typhus, plague and tapeworm. Inaddition, fleas are a problem for animals maintained as pets because theinfestation becomes a source of annoyance for the pet owner who may findhis or her home generally contaminated with fleas which feed on thepets. As such, fleas are a problem not only when they are on an animalbut also when they are in the general environment of the animal.

The medical and veterinary importance of flea infestation has promptedthe development of reagents capable of controlling flea infestation.Commonly encountered methods to control flea infestation are generallyfocussed on use of insecticides in formulations such as sprays,shampoos, dusts, dips, or foams, or in pet collars. While some of theseproducts are efficacious, most, at best, offer protection of a verylimited duration. Furthermore, many of the methods are often notsuccessful in reducing flea populations on the pet for one or more ofthe following reasons: (1) failure of owner compliance (frequentadministration is required); (2) behavioral or physiological intoleranceof the pet to the pesticide product or means of administration; and (3)the emergence of flea populations resistant to the prescribed dose ofpesticide. Additional anti-flea products include nontoxic reagents suchas insect growth regulators (IGRs), including methoprene, which mimicsflea hormones and affect flea larval development.

An alternative method for controlling flea infestation is the use offlea vaccines to be administered to animals prior to or during fleainfestation. However, despite considerable interest in developinganti-flea reagents, no flea vaccine presently exists.

SUMMARY OF THE INVENTION

The present invention relates to flea serine protease proteins, to fleaaminopeptidase proteins, and to flea cysteine protease proteins; to fleaserine protease, aminopeptidase and/or cysteine protease nucleic acidmolecules, including those that encode such proteins; to antibodiesraised against such proteins; and to compounds that inhibit flea serineprotease, aminopeptidase and/or cysteine protease activities. Thepresent invention also includes methods to obtain such proteins, nucleicacid molecules, antibodies, and inhibitors. Also included in the presentinvention are therapeutic compositions comprising such proteins, nucleicacid molecules, antibodies, and/or inhibitors as well as the use of suchtherapeutic compositions to protect a host animal from flea infestation.

One embodiment of the present invention is an isolated flea serineprotease nucleic acid molecule that hybridizes under stringenthybridization conditions with a flea serine protease gene. Particularlypreferred flea serine protease nucleic acid molecules include nucleicacid sequences SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22,SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32,SEQ ID NO:34, SEQ ID NO:36, and SEQ ID NO:38, SEQ ID NO:52, SEQ IDNO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ IDNO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ IDNO:74, SEQ ID NO:76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82, SEQ IDNO:84, SEQ ID NO:86, SEQ ID NO:88, SEQ ID NO:90, SEQ ID NO:92, SEQ IDNO:94, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102, SEQ IDNO:104, SEQ ID NO:106, SEQ ID NO:108, SEQ ID NO:110, the nucleic acidsequences disclosed in Table 2, SEQ ID NO:120, SEQ ID NO:122, SEQ IDNO:124, SEQ ID NO:126, SEQ ID NO:128, SEQ ID NO:130, SEQ ID NO:132, SEQID NO:134, SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142,SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ ID NO:150, SEQ IDNO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:158, SEQ ID NO:160, SEQID NO:162, SEQ ID NO:164 and/or SEQ ID NO:189 and/or nucleic acidsequences encoding proteins having amino acid sequences SEQ ID NO:1, SEQID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:17, SEQ ID NO:19, SEQ IDNO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ IDNO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ IDNO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ IDNO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:55, SEQ IDNO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ IDNO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ IDNO:77, SEQ ID NO:79, SEQ ID NO:81, SEQ ID NO:83, SEQ ID NO:85, SEQ IDNO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:93, SEQ ID NO:95, SEQ IDNO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, SEQ IDNO:107, SEQ ID NO:109, SEQ ID NO:111, SEQ ID NO:121, SEQ ID NO:123, SEQID NO:125, SEQ ID NO:127, SEQ ID NO:129, SEQ ID NO:131, SEQ ID NO:133,SEQ ID NO:135, SEQ ID NO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ IDNO:143, SEQ ID NO:145, SEQ ID NO:147, SEQ ID NO:149, SEQ ID NO:151, SEQID NO:153, SEQ ID NO:155, SEQ ID NO:157, SEQ ID NO:159, SEQ ID NO:161,SEQ ID NO:163, SEQ ID NO:165, SEQ ID NO:179, SEQ ID NO:180, SEQ IDNO:181, SEQ ID NO:184 and/or SEQ ID NO:190, and/or the amino acidsequences disclosed in Table 2; as well as allelic variants of any ofthe listed nucleic acid sequences or complements of any of the listednucleic acid sequences.

Another embodiment of the present invention is an isolated fleaaminopeptidase nucleic acid molecule that hybridizes under stringenthybridization conditions with a flea aminopeptidase gene. A particularlypreferred flea aminopeptidase nucleic acid molecule includes nucleicacid sequence SEQ ID NO:50, SEQ ID NO:112, SEQ ID NO:169 and/or SEQ IDNO:171, and/or nucleic acid sequences encoding proteins having aminoacid sequences SEQ ID NO:51, SEQ ID NO:113, SEQ ID NO:167, SEQ IDNO:170, SEQ ID NO:172, as well as allelic variants of any of thosenucleic acid sequences or complements of any of the listed nucleic acidsequences.

Another embodiment of the present invention is an isolated flea cysteineprotease nucleic acid molecule that hybridizes under stringenthybridization conditions with a flea cysteine protease gene. Aparticularly preferred flea cysteine protease nucleic acid moleculeincludes nucleic acid sequence SEQ ID NO:177, and/or nucleic acidsequences encoding proteins having amino acid sequence SEQ ID NO:178, aswell as allelic variants of any of those nucleic acid sequences orcomplements of any of the listed nucleic acid sequences.

The present invention also includes a nucleic acid molecule comprising anucleic acid sequence that encodes a flea protease protein, including alarval serine protease protein, an adult serine protease protein, alarval aminopeptidase protein, an adult aminopeptidase protein, a larvalcysteine protease protein and/or an adult cysteine protease protein

The present invention also relates to recombinant molecules, recombinantviruses and recombinant cells that include flea serine protease,aminopeptidase and/or cysteine protease nucleic acid molecules of thepresent invention. Also included are methods to produce such nucleicacid molecules, recombinant molecules, recombinant viruses andrecombinant cells.

Another embodiment of the present invention includes an isolated fleaserine protease protein, including a protein that includes a flea serineprotease protein. A preferred flea serine protease protein is capable ofeliciting an immune response against a natural flea protease whenadministered to an animal and/or of having serine protease activity.Particularly preferred flea serine protease proteins are those encodedby preferred flea serine protease nucleic acid molecules of the presentinvention.

Yet another embodiment of the present invention includes an isolatedflea aminopeptidase protein, including a protein that includes a fleaaminopeptidase protein. A preferred flea aminopeptidase protein iscapable of eliciting an immune response against a natural flea proteasewhen administered to an animal and/or of having aminopeptidase activity.

Yet another embodiment of the present invention includes an isolatedflea cysteine protease protein, including a protein that includes a fleacysteine protease protein. A preferred flea cysteine protease protein iscapable of eliciting an immune response against a natural flea proteasewhen administered to an animal and/or of having cysteine proteaseactivity.

The present invention also relates to mimetopes of flea serine protease,aminopeptidase and/or cysteine protease proteins as well as to isolatedantibodies that selectively bind to flea serine protease proteins ormimetopes thereof, to flea aminopeptidase proteins or mimetopes thereof,or to flea cysteine protease proteins or mimetopes thereof. Alsoincluded are methods, including recombinant methods, to produceproteins, mimetopes and antibodies of the present invention.

Yet another embodiment of the present invention is a therapeuticcomposition that is capable of reducing flea infestation. Such atherapeutic composition includes one or more of the following protectivecompounds: an isolated flea serine protease protein or a mimetopethereof; an isolated flea serine protease nucleic acid molecule thathybridizes under stringent hybridization conditions with a flea serineprotease gene; an isolated antibody that selectively binds to a fleaserine protease protein; an inhibitor of flea serine protease activityidentified by its ability to inhibit flea serine protease activity; anisolated flea aminopeptidase protein or a mimetope thereof; an isolatedflea aminopeptidase nucleic acid molecule that hybridizes understringent hybridization conditions with a flea aminopeptidase gene; anisolated antibody that selectively binds to a flea aminopeptidaseprotein; an inhibitor of flea aminopeptidase activity identified by itsability to inhibit flea aminopeptidase activity; an isolated fleacysteine protease protein or a mimetope thereof; an isolated fleacysteine protease nucleic acid molecule that hybridizes under stringenthybridization conditions with a flea cysteine protease gene; an isolatedantibody that selectively binds to a flea cysteine protease protein; aninhibitor of flea cysteine protease activity identified by its abilityto inhibit flea cysteine protease activity. A preferred therapeuticcomposition of the present invention also includes an excipient, anadjuvant and/or a carrier. Also included in the present invention is amethod to reduce flea infestation. The method includes the step ofadministering to the animal a therapeutic composition of the presentinvention.

Another embodiment of the present invention is a method to identify acompound capable of inhibiting flea serine protease, flea aminopeptidaseor flea cysteine protease activity. The method includes the steps of:(a) contacting an isolated flea serine protease protein, a fleaaminopeptidase protein or a flea cysteine protease protein, with aputative inhibitory compound under conditions in which, in the absenceof the compound, the protein has, respectively, serine protease,aminopeptidase or cysteine protease activity; and (b) determining if theputative inhibitory compound inhibits the respective activity. Alsoincluded in the present invention is a test kit to identify a compoundcapable of inhibiting flea serine protease, flea aminopeptidase orcysteine protease activity. Such a kit includes an isolated flea serineprotease protein having serine protease activity, an isolated fleaaminopeptidase protein having aminopeptidase activity or an isolatedflea cysteine protease protein having cysteine protease activity and ameans for determining the extent of inhibition of the respectiveactivity in the presence of a putative inhibitory compound.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts a protease substrate gel analysis of the relativeproteolytic activity in 1, 2, 5 or 10 midguts from either fed or unfedfemale fleas.

FIG. 2 depicts a protease substrate gel analysis of fed and unfed midgutpreparations incubated in the presence or absence of a serine proteaseinhibitor.

FIG. 3 depicts a protease substrate gel analysis of various fractionsobtained in the preparation of a soluble flea midgut preparationincubated in the presence or absence of a serine protease inhibitor.

FIG. 4 depicts a protease substrate gel showing midgut protease activityas a function of time after flea blood feeding.

FIG. 5A depicts a Coomassie stained SDS-PAGE of partially purified(1,3-³ H)-diisopropylfluoro-phosphate (DFP)-labeled fed flea midgutserine proteases.

FIG. 5B depicts an autoradiogram of the SDS-PAGE gel of FIG. 5A ofpartially purified DFP-labeled fed flea midgut serine proteases.

FIG. 6 depicts the mean viability of adult (both male and female) fleasfed blood containing certain protease inhibitors.

FIG. 7 depicts the mean fecundity of adult female fleas fed bloodcontaining certain protease inhibitors.

FIG. 8 depicts the mean viability of adult (both male and female) fleasfed blood containing certain protease inhibitors.

FIG. 9 depicts the mean fecundity of adult female fleas fed bloodcontaining certain protease inhibitors.

FIG. 10 depicts induction over time of DFP-labeled proteases in fleasfeeding on a cat.

FIG. 11 depicts SDS-PAGE of induction over time of DFP-labeled proteasesin fleas feeding on a cat.

FIG. 12 depicts SDS-PAGE of DFP-labeled larval proteases in unfedlarvae, fed 1st instar larvae and fed 3rd instar larvae.

DETAILED DESCRIPTION OF THE INVENTION

The present invention includes the use of compounds that inhibit fleaprotease activity to protect a host animal from flea infestation. Theinventors have discovered that proteases are significant components ofthe flea midgut and are good targets for immunotherapeutic and/orchemotherapeutic intervention to reduce flea burden both on the hostanimal and in the immediate (i.e., surrounding) environment of theanimal. The inventors have shown, for example, that the viability and/orfecundity of fleas consuming a blood meal is reduced when the blood mealcontains compounds that reduce flea protease activity, probably becausethe compounds interfere with flea digestion and other functions.Compounds that reduce the amount and/or activity of flea proteaseswithout substantially harming the host animal are included in thepresent invention. Such compounds include flea protease vaccines,anti-flea protease antibodies, flea protease inhibitors, and/orcompounds that suppress protease synthesis; such compounds are discussedin more detail below.

One embodiment of the present invention is a method to protect a hostanimal from flea infestation by treating the animal with a compositionthat includes a compound that reduces the protease activity of fleasfeeding (includes fleas in the process of feeding as well as fleashaving fed) from the treated animal thereby reducing the flea burden onthe animal and in the environment of the animal. It is to be noted thatthe term "a" or "an" entity refers to one or more of that entity; forexample, a compound refers to one or more compounds. As such, the terms"a" (or "an"), "one or more" and "at least one" can be usedinterchangeably herein. Thus, a composition of the present invention caninclude one or more compounds that target (reduced the activity of) oneor more proteases in the flea.

As used herein, the phrase "to protect an animal from flea infestation"refers to reducing the potential for flea population expansion on andaround the animal (i.e., reducing the flea burden). Preferably, the fleapopulation size is decreased, optimally to an extent that the animal isno longer bothered by fleas. A host animal, as used herein, is an animalfrom which fleas can feed by attaching to and feeding through the skinof the animal. Fleas, and other ectoparasites, can live on a host animalfor an extended period of time or can attach temporarily to an animal inorder to feed. At any given time, a certain percentage of a fleapopulation can be on a host animal whereas the remainder can be in theenvironment surrounding the animal (i.e., in the environment of theanimal). Such an environment can include not only adult fleas, but alsoflea eggs and/or flea larvae. The environment can be of any size suchthat fleas in the environment are able to jump onto and off of a hostanimal. As such, it is desirable not only to reduce the flea burden onan animal per se, but also to reduce the flea burden in the environmentsurrounding the animal.

In accordance with the present invention, a host animal is treated byadministering to the animal a compound of the present invention in sucha manner that the compound itself (e.g., a protease inhibitor, proteasesynthesis suppressor or anti-flea protease antibody) or a productgenerated by the animal in response to administration of the compound(e.g., antibodies produced in response to a flea protease vaccine, orconversion of an inactive inhibitor "prodrug" to an active proteaseinhibitor) ultimately enters the flea midgut. An animal is preferablytreated in such a way that the compound or product thereof enters theblood stream of the animal. Fleas are then exposed to the compound whenthey feed from the animal. For example, flea protease inhibitorsadministered to an animal are administered in such a way that theinhibitors enter the blood stream of the animal, where they can be takenup by feeding fleas. In another embodiment, when a host animal isadministered a flea protease vaccine, the treated animal mounts animmune response resulting in the production of antibodies against theprotease (anti-flea protease antibodies) which circulate in the animal'sblood stream and are taken up by fleas upon feeding. Blood taken up byfleas enters the flea midgut where compounds of the present invention,or products thereof, such as anti-flea protease antibodies, fleaprotease inhibitors, and/or protease synthesis suppressors, interactwith, and reduce proteolytic activity in the flea midgut. The presentinvention also includes the ability to reduce larval flea infestation inthat when fleas feed from a host animal that has been administered atherapeutic composition of the present invention, at least a portion ofcompounds of the present invention, or products thereof, in the bloodtaken up by the flea are excreted by the flea in feces, which issubsequently ingested by flea larvae. It is of note that flea larvaeobtain most, if not all, of their nutrition from flea feces.

In accordance with the present invention, reducing proteolytic activityin flea midguts can lead to a number of outcomes that reduce flea burdenon treated animals and their surrounding environments. Such outcomesinclude, but are not limited to, (a) reducing the viability of fleasthat feed from the treated animal, (b) reducing the fecundity of femalefleas that feed from the treated animal, (c) reducing the reproductivecapacity of male fleas that feed from the treated animal, (d) reducingthe viability of eggs laid by female fleas that feed from the treatedanimal, (e) altering the blood feeding behavior of fleas that feed fromthe treated animal (e.g., fleas take up less volume per feeding or feedless frequently), (f) reducing the viability of flea larvae, for exampledue to the feeding of larvae from feces of fleas that feed from thetreated animal and/or (g) altering the development of flea larvae (e.g.,by decreasing feeding behavior, inhibiting growth, inhibiting (e.g.,slowing or blocking) molting, and/or otherwise inhibiting maturation toadults). One embodiment of the present invention is a composition thatincludes one or more compounds that reduce the activity of one or moreflea proteases directly (e.g., an anti-flea protease antibody or a fleaprotease inhibitor) and/or indirectly (e.g., a flea protease vaccine).Suitable flea proteases to target include flea aminopeptidases, fleacarboxypeptidases and/or flea endopeptidases. Such proteases can includecytosolic and/or membrane-bound forms of a protease. Preferred fleaproteases to target include, but are not limited to, serine proteases,metalloproteases, aspartic acid proteases and/or cysteine proteases. Itis to be noted that these preferred groups of proteases includeaminopeptidases, carboxypeptidases and/or endopeptidases. Preferred fleaproteases to target include, but are not limited to, proteases thatdegrade hemoglobin, proteases involved in blood coagulation and/or lytic(anti-coagulation) pathways, proteases involved in the maturation ofpeptide hormones, proteases that inhibit complement or other host immuneresponse elements (e.g., antibodies) and/or proteases involved invitellogenesis. A number of proteases are known to those skilled in theart, including, but not limited to, aminopeptidases, such as leucineaminopeptidase and aminopeptidases B and M; astacin-likemetalloproteases; calpains; carboxypeptidases, such as carboxypeptidasesA, P and Y; cathepsins, such as cathepsins B, D, E, G, H, and L,chymotrypsins; cruzipains; meprins; papains; pepsins; renins;thermolysins and trypsins. A particularly preferred protease to targetis a protease having a proteolytic activity that, when targeted with acomposition of the present invention, reduces flea burden withoutsubstantially harming the host animal. Such a protease can be identifiedusing, for example, methods as disclosed herein.

One aspect of the present invention is the discovery that a substantialamount of the proteolytic activity found in flea midguts is serineprotease activity. Both in vitro and in vivo studies using a number ofprotease inhibitors substantiate this discovery, details of which aredisclosed in the Examples. As such a particularly preferred protease totarget is a serine protease. Examples of serine proteases, include, butare not limited to, acrosins, bromelains, cathepsin G, chymotrypsins,collagenases, elastases, factor Xa, ficins, kallikreins, papains,plasmins, Staphylococcal V8 proteases, thrombins and trypsins. In oneembodiment, a preferred flea serine protease to target includes aprotease having trypsin-like or chymotrypsin-like activity. It isappreciated by those skilled in the art that an enzyme having "like"proteolytic activity has similar activity to the referenced protease,although the exact structure of the preferred substrate cleaved maydiffer. "Like" proteases usually have similar tertiary structures astheir referenced counterparts.

Protease inhibitor studies disclosed in the Examples section alsoindicate that additional preferred proteases to target includeaminopeptidases and/or metalloproteases. Examples of such proteasesinclude exo- and endo-metalloproteases, digestive enzymes, and enzymesinvolved in peptide hormone maturation. One example of an aminopeptidasethat is also a metalloprotease is leucine aminopeptidase.

Suitable compounds to include in compositions of the present inventioninclude, but are not limited to, a vaccine comprising a flea protease (aflea protease vaccine), an antibody that selectively binds to a fleaprotease (an anti-flea protease antibody), a flea protease inhibitor (acompound other than a vaccine or an antibody that inhibits a fleaprotease), and a mixture of such compounds. As used herein, a mixturethereof refers to a combination of one or more of the cited entities.Compositions of the present invention can also include compounds tosuppress protease synthesis or maturation, such as, but not limited to,protease modulating peptides.

A preferred embodiment of the present invention is a flea proteasevaccine and its use to reduce the flea population on and around ananimal. A flea protease vaccine can include one or more proteins capableof eliciting an immune response against a flea protease and can alsoinclude other components. Preferred flea protease vaccines include aflea serine protease, a flea metalloprotease, a flea aspartic acidprotease and/or a flea cysteine protease, with flea serine protease,flea metalloprotease and/or flea aminopeptidase vaccines being morepreferred. Examples of flea protease vaccines include soluble fleamidgut preparations of the present invention as well as one or moreisolated proteins of the present invention.

One embodiment of the present invention is a soluble flea midgutpreparation. Such a preparation includes primarily components naturallypresent in the lumen of a flea midgut and, depending on the method ofpreparation, can also include one or more peripheral midgut membraneproteins. Methods to preferentially include, or exclude, membraneproteins from such a preparation are known to those skilled in the art.The present invention includes the discovery that such a preparation hasproteolytic activity, of which a substantial portion is serine proteaseactivity. Preferably at least about 70 percent of the proteolyticactivity in a soluble flea midgut soluble preparation is serine proteaseactivity, as can be indicated by the ability to inhibit at least about70 percent of the proteolytic activity with4-2-aminoethyl-benzenesulfonylfluoride-hydrochloride (AEBSF). Serineprotease activity can also be identified using other known inhibitors orsubstrates. Other preferred inhibitors that can inhibit at least about70 percent of the proteolytic activity of a soluble flea midgutpreparation of the present invention include soybean trypsin inhibitor,1,3-diisopropylfluorophosphate or leupeptin.

A soluble flea midgut preparation of the present invention includesproteases that range in molecular weight from about 5 kilodaltons (kD orkDa) to about 200 kD, as determined by SDS-PAGE (sodium dodecyl sulfatepolyacrylamide gel electrophoresis), with at least a substantial portionof the serine proteases ranging in molecular weight from about 5 kD toabout 60 kD, as determined by SDS-PAGE. A substantial portion ofprotease activity in a soluble flea midgut preparation of the presentinvention has a pH activity optimum ranging from about pH 5 to about pH10, preferably an activity optimum ranging from about pH 7 to about pH9, and even more preferably an activity optimum of about pH 8. While notbeing bound by theory, such a pH optimum suggests that a largeproportion of proteases in soluble flea midgut preparations of thepresent invention are serine proteases. It is also interesting to notethat the pH of the flea midgut is also about pH 8. The findings thatproteases in soluble flea midgut preparations of the present inventionexhibit a varied pattern of inhibition by protease inhibitors of a giventype (e.g., serine protease inhibitors), as well as variances seen inmolecular weights and pH optima of the proteases, suggest that there area number of protease isoforms in such preparations.

A soluble flea midgut preparation of the present invention is preferablyprepared by a method that includes the steps of (a) disrupting a fleamidgut to produce a mixture including a liquid portion and a solidportion and (b) recovering the liquid portion to obtain a soluble fleamidgut preparation. Such a method is a simplified version of methodsdisclosed in U.S. Pat. No. 5,356,622, ibid. It is to be noted that inaccordance with the present invention, methods disclosed in U.S. Pat.No. 5,356,622, ibid. can also be used to prepare soluble flea midgutpreparations having similar proteolytic activities.

Flea midguts can be obtained (e.g., dissected from) from unfed fleas orfrom fleas that recently consumed a blood meal (i.e., blood-fed fleas).Such midguts are referred to herein as, respectively, unfed flea midgutsand fed flea midguts. Flea midguts can be obtained from either male orfemale fleas. As demonstrated in the Examples section, female fleamidguts exhibit somewhat more proteolytic activity than do male fleamidguts. Furthermore, fed flea midguts have significantly moreproteolytic activity than do unfed flea midguts. While not being boundby theory, it is believed that blood feeding induces in flea midguts thesynthesis and/or activation of proteases as well as other factors (e.g.,enzymes, other proteins, co-factors, etc.) important in digesting theblood meal, as well as in neutralizing host molecules potentiallydamaging to the flea (e.g., complement, immunoglobulins, bloodcoagulation factors). It is also to be appreciated that unfed fleamidguts may contain significant targets not found in fed flea midgutsand vice versa. Furthermore, although the present application focussesprimarily on flea midgut proteases, it is to be noted that the presentinvention also includes other components of soluble flea midgutpreparations of the present invention that provide suitable targets toreduce flea burden on an animal and in the environment of that animal;see also U.S. Pat. No. 5,356,622, ibid.

Methods to disrupt flea midguts in order to obtain a soluble flea midgutpreparation are known to those skilled in the art and can be selectedaccording to, for example, the volume being processed and the buffersbeing used. Such methods include any technique that promotes cell lysis,such as, but are not limited to, chemical disruption techniques (e.g.,exposure of midguts to a detergent) as well as mechanical disruptiontechniques (e.g., homogenization, sonication, use of a tissue blender orglass beads, and freeze/thaw techniques).

Methods to recover a soluble flea midgut preparation are also known tothose skilled in the art and can include any method by which the liquidportion of disrupted flea midguts is separated from the solid portion(e.g., filtration or centrifugation). In a preferred embodiment,disrupted flea midguts are subjected to centrifugation, preferably at anacceleration ranging from about 10,000×g to about 15,000×g for severalminutes (e.g., from about 1 min. to about 15 min.). The supernatant fromsuch a centrifugation comprises a soluble flea midgut preparation of thepresent invention.

The present invention also includes an isolated protein that includes anamino acid sequence encoded by a nucleic acid molecule capable ofhybridizing under stringent conditions (i.e., that hybridize understringent hybridization conditions) with a nucleic acid molecule thatencodes a protease present (i.e., the nucleic acid molecules hybridizewith the nucleic acid strand that is complementary to the coding strand)in (i.e., can be found in) a flea midgut, such as a midgut from ablood-fed female flea, a midgut from a blood-fed male flea, a midgutfrom an unfed female flea or a midgut from an unfed male flea. Apreferred midgut protease is present in the lumen of the midgut.

An isolated protein of the present invention, also referred to herein asan isolated protease protein, preferably is capable of eliciting animmune response against a flea midgut protease and/or has proteolyticactivity. According to the present invention, an isolated, orbiologically pure, protein, is a protein that has been removed from itsnatural milieu. As such, "isolated" and "biologically pure" do notnecessarily reflect the extent to which the protein has been purified.An isolated protease protein can be obtained from its natural source.Such an isolated protein can also be produced using recombinant DNAtechnology or chemical synthesis.

As used herein, an isolated protein of the present invention can be afull-length protein or any homologue of such a protein, such as aprotein in which amino acids have been deleted (e.g., a truncatedversion of the protein, such as a peptide), inserted, inverted,substituted and/or derivatized (e.g., by glycosylation, phosphorylation,acetylation, myristoylation, prenylation, palmitoylation, amidationand/or addition of glycerophosphatidyl inositol) such that the homologuecomprises a protein having an amino acid sequence that is sufficientlysimilar to a natural flea midgut protease that a nucleic acid sequenceencoding the homologue is capable of hybridizing under stringentconditions to (i.e., with) the complement of a nucleic acid sequenceencoding the corresponding natural flea midgut protease amino acidsequence. As used herein, stringent hybridization conditions refer tostandard hybridization conditions under which nucleic acid molecules,including oligonucleotides, are used to identify similar nucleic acidmolecules. Such standard conditions are disclosed, for example, inSambrook et al., Molecular Cloning: A Laboratory Manual, Cold SpringHarbor Labs Press, 1989; Sambrook et al., ibid., is incorporated byreference herein in its entirety. Stringent hybridization conditionstypically permit isolation of nucleic acid molecules having at leastabout 70% nucleic acid sequence identity with the nucleic acid moleculebeing used to probe in the hybridization reaction. Formulae to calculatethe appropriate hybridization and wash conditions to achievehybridization permitting 30% or less mismatch of nucleotides aredisclosed, for example, in Meinkoth et al., 1984, Anal. Biochem. 138,267-284; Meinkoth et al., ibid., is incorporated by reference herein inits entirety.

The minimal size of a protein homologue of the present invention is asize sufficient to be encoded by a nucleic acid molecule capable offorming a stable hybrid with the complementary sequence of a nucleicacid molecule encoding the corresponding natural protein. As such, thesize of the nucleic acid molecule encoding such a protein homologue isdependent on nucleic acid composition and percent homology between thenucleic acid molecule and complementary sequence as well as uponhybridization conditions per se (e.g., temperature, salt concentration,and formamide concentration). The minimal size of such nucleic acidmolecules is typically at least about 12 to about 15 nucleotides inlength if the nucleic acid molecules are GC-rich and at least about 15to about 17 bases in length if they are AT-rich. As such, the minimalsize of a nucleic acid molecule used to encode a protease proteinhomologue of the present invention is from about 12 to about 18nucleotides in length. There is no limit, other than a practical limit,on the maximal size of such a nucleic acid molecule in that the nucleicacid molecule can include a portion of a gene, an entire gene, ormultiple genes, or portions thereof. Similarly, the minimal size of aprotease protein homologue of the present invention is from about 4 toabout 6 amino acids in length, with preferred sizes depending on whethera full-length, multivalent (i.e., fusion protein having more than onedomain each of which has a function), or functional portions of suchproteins are desired. Protease protein homologues of the presentinvention preferably have protease activity and/or are capable ofeliciting an immune response against a flea midgut protease.

A protease protein homologue of the present invention can be the resultof allelic variation of a natural gene encoding a flea protease. Anatural gene refers to the form of the gene found most often in nature.Protease protein homologues can be produced using techniques known inthe art including, but not limited to, direct modifications to a geneencoding a protein using, for example, classic or recombinant DNAtechniques to effect random or targeted mutagenesis. Isolated proteaseproteins of the present invention, including homologues, can beidentified in a straight-forward manner by the proteins' ability toeffect proteolytic activity and/or to elicit an immune response againsta flea midgut protease. Such techniques are known to those skilled inthe art.

A preferred protease protein of the present invention is a flea serineprotease, a flea metalloprotease, a flea aspartic acid protease, a fleacysteine protease, or a homologue of any of these proteases. A morepreferred protease protein is a flea serine protease, a fleametalloprotease or a homologue of either. Also preferred is a fleaaminopeptidase or a homologue thereof. Also preferred is a flea cysteineprotease or a homologue thereof. Particularly preferred is a flea serineprotease or a homologue thereof.

Preferred protease proteins of the present invention are flea proteaseproteins having molecular weights ranging from about 5 kD to about 200kD, as determined by SDS-PAGE, and homologues of such proteins. Morepreferred are flea protease proteins having molecular weights rangingfrom about 5 kD to about 60 kD, as determined by SDS-PAGE, andhomologues of such proteins. Even more preferred are flea serineprotease proteins, particularly those having molecular weights of about26 kD (denoted PfSP26, now denoted PafSP-26K to distinguish from fleaPfSP26 as described in Example 26), about 24 kD (denoted PfSP24, nowdenoted PafSP-24K to distinguish from flea PfSP24 as described inExample 27), about 19 kD (denoted PfSP19,now denoted PafSP-19K todistinguish from flea PfSP19 as described in Example 32), about 6 kD(denoted PfSP6, now denoted PafSP-6K to distinguish from flea PfSP6 asdescribed in Example 11), about 31 kD (denoted PfSP28), about 25 kD(denoted PlfSP-25K1) from 1st instar larvae, about 25 kD (denotedPlfSP-25K3) from 3rd instar larvae, about 28 kD (denoted PlfSP-28K3) andabout 31 kD (denoted PlSP-31K3), and flea aminopeptidase proteins,particularly those having molecular weights of about 95 kD (denotedPfAP-95K) as determined by SDS-PAGE, and homologues of such proteins.

One preferred embodiment of the present invention is an isolated fleaprotease protein that includes an amino acid sequence encoded by anucleic acid molecule that hybridizes under stringent hybridizationconditions with a flea serine protease gene, with a flea aminopeptidasegene or with a flea cysteine protease gene. As used herein, a fleaprotease gene includes all nucleic acid sequences related to a naturalflea protease gene such as regulatory regions that control production ofa flea protease protein encoded by that gene (such as, but not limitedto, transcription, translation or post-translation control regions) aswell as the coding region itself.

The inventors have discovered an extensive family of serine proteases,encoded by a family of serine protease genes. Such a gene family may bedue to allelic variants (i.e., genes having similar, but different,sequences at a given locus in a population of fleas) and/or to, theexistence of serine protease genes at more than one locus in the fleagenome. As such, the present invention includes flea serine proteasegenes comprising not only the nucleic acid sequences disclosed herein(e.g., genes including nucleic acid sequences SEQ ID NO:16, SEQ IDNO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ IDNO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ IDNO:38 and/or the nucleic acid sequences disclosed in Table 2 (i.e., SEQID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ IDNO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ IDNO:72, SEQ ID NO:74, SEQ ID NO:76, SEQ ID NO:78), SEQ ID NO:80, SEQ IDNO:82, SEQ ID NO:84, SEQ ID NO:86, SEQ ID NO:88, SEQ ID NO:90, SEQ IDNO:92, SEQ ID NO:94, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ IDNO:102, SEQ ID NO:104, SEQ ID NO:106, SEQ ID NO:108, SEQ ID NO:110, SEQID NO:120, SEQ ID NO:122, SEQ ID NO:124, SEQ ID NO:126, SEQ ID NO:128,SEQ ID NO:130, SEQ ID NO:132, SEQ ID NO:134, SEQ ID NO:136, SEQ IDNO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQID NO:148, SEQ ID NO:150, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156,SEQ ID NO:158, SEQ ID NO:160, SEQ ID NO:162, SEQ ID NO:164 and/or SEQ IDNO:189, and/or nucleic acid sequences encoding proteins having aminoacid sequences as disclosed herein (e.g., SEQ ID NO:1, SEQ ID NO:2, SEQID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ IDNO:8, SEQ ID NO:9, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ IDNO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ IDNO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:40, SEQ IDNO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ IDNO:46, SEQ ID NO:47, the amino acid sequences disclosed in Table 2(i.e., SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ IDNO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ IDNO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79), SEQ IDNO:81, SEQ ID NO:83, SEQ ID NO:85, SEQ ID NO:87, SEQ ID NO:89, SEQ IDNO:91, SEQ ID NO:93, SEQ ID NO:95, SEQ ID NO:97, SEQ ID NO:99, SEQ IDNO:101, SEQ ID NO:103, SEQ ID NO:105, SEQ ID NO:107, SEQ ID NO:109, SEQID NO:111, SEQ ID NO:121, SEQ ID NO:123, SEQ ID NO:125, SEQ ID NO:127,SEQ ID NO:129, SEQ ID NO:131, SEQ ID NO:133, SEQ ID NO:135, SEQ IDNO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ ID NO:143, SEQ ID NO:145, SEQID NO:147, SEQ ID NO:149, SEQ ID NO:151, SEQ ID NO:153, SEQ ID NO:155,SEQ ID NO:157, SEQ ID NO:159, SEQ ID NO:161, SEQ ID NO:163, SEQ IDNO:165, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:184and/or SEQ ID NO:190), but also allelic variants of any of those nucleicacid sequences. (It should be noted that since nucleic acid sequencingtechnology is not entirely error-free, all sequences represented hereinare at best apparent (i.e., deduced) nucleic acid or amino acidsequences.)

A preferred flea aminopeptidase gene includes nucleic acid sequence SEQID NO:50, which encodes an aminopeptidase protein including SEQ IDNO:51. Another preferred flea aminopeptidase gene includes nucleic acidsequence SEQ ID NO:112, which encodes an aminopeptidase proteinincluding SEQ ID NO:113. Another preferred flea aminopeptidase geneincludes nucleic acid sequences SEQ ID NO:169 and/or SEQ ID NO:171; thegene encodes a putative membrane-bound form of an aminopeptidase proteinthat includes SEQ ID NO:167, SEQ ID NO:170 and/or SEQ ID NO:172.Additional preferred aminopeptidase genes include allelic variants ofSEQ ID NO:50, of SEQ ID NO:112 SEQ ID NO:169 or SEQ ID NO:171.

A preferred flea cysteine protease gene includes nucleic acid sequenceSEQ ID NO:SEQ ID NO:177, which encodes a cysteine protease proteinincluding SEQ ID NO:178. Additional preferred cysteine protease genesinclude allelic variants of SEQ ID NO:177.

A preferred flea serine protease protein of the present invention isencoded by a nucleic acid molecule that hybridizes under stringenthybridization conditions with at least one of the following nucleic acidmolecules: nfSP1, nfSP2, nfSP3, nfSP4, nfSP5, nfSP6, nfSP7, nfSP8,nfSP9, nfSP10, nfSP11, nfSP12, nfSP13, nfSP14, nfSP15, nfSP16, nfSP17,nfSP18, nfSP19, nfSP20, nfSP21, nfSP23, nfSP24, nfSP25, nfSP26, nfSP27,nfSP28, nfSP29, nfSP30, nfSP31, nfSP32, nfSP33, nfSP34, nfSP36, nfSP37,nfSP38 and nfSP39. As used herein, each of these nucleic acid moleculesrepresent the entire coding region of a flea serine protease gene of thepresent invention (at least portions of which are also referred to byflea clone numbers, as described in the Examples). Nucleic acidmolecules that contain partial coding regions or other parts of thecorresponding gene are denoted by names that include the size of thosenucleic acid molecules (e.g., nfSP4₁₅₆). Nucleic acid moleculescontaining apparent full length coding regions for which the size isknown also are denoted by names that include the size of those nucleicacid molecules (e.g., nfSP4₆₇₂). The production, and at least partialnucleic acid sequence, of such nucleic acid molecules is disclosed inthe Examples.

Particularly preferred serine protease proteins are encoded by a nucleicacid molecule that hybridizes under stringent hybridization conditionswith at least one of the following nucleic acid molecules: nfSP4₆₇₂,nfSP1₁₅₆, nfSP2₁₆₈, nfSP3₁₇₇, nfSP4₁₅₆, nfSP5₁₅₉, nfSP6₁₆₈, nfSP7₁₅₉,nfSP8₁₈₆, nfSP9₁₆₈, nfSP10₁₂₀, and nfSP11₁₆₂ as well as other specificnucleic acid molecules disclosed in the Examples section, such as, butnot limited to, nfSP1₇₇₉, nfSP2₉₄₄, nfSP3₁₇₇, nfSP4₆₇₂, nfSP5₁₅₇,nfSP5₂₁₈, nfSP6₉₃₂, nfSP7₈₉₄, nfSP8₂₉₉, nfSP9₂₆₆, nfSP10₃₇₈, nfSP11₂₅₂,nfSP12₁₄₄, nfSP12₂₂₅, nfSP13₈₅₀, nfSP14₂₁₃, nfSP15₂₅₂, nfSP16₁₆₈,nfSP18₅₃₄, nfSP19₃₅₉, nfSP20₈₄₁, nfSP5₈₀₆, nfSP11₃₀₇, nfSP8₅₁₅,nfSP12₇₅₈, nfSP26₆₁₀, nfSP27₃₈₆, nfSP23₄₂₃, nfSP24₄₁₀, nfSP33 426,nfSP34₃₉₀, nfSP36₁₉₇, nfSP38₃₄₁, nfSP37₂₆₁, nfSP39₂₆₇ nfSP29₆₁₂,nfSP30₆₄₁, nfSP31₆₂₆, nfSP32₄₃₃, nfSP15₈₁₅, nfSP19₈₅₅, nfSP25₈₆₄,nfSP21₅₉₅, nfSP28₉₂₃ and/or nfSP33₇₇₈. Even more preferred serineprotease proteins include the following amino acid sequences: SEQ IDNO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6,SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:17, SEQ ID NO:19, SEQID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ IDNO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ IDNO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ IDNO:45, SEQ ID NO:46, SEQ ID NO:47, and/or the amino acid sequencespresented in Table 2 (i.e., SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57,SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67,SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77,and/or SEQ ID NO:79), as well as SEQ ID NO:81, SEQ ID NO:83, SEQ IDNO:85, SEQ ID NO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:93, SEQ IDNO:95, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ IDNO:105, SEQ ID NO:107, SEQ ID NO:109, SEQ ID NO:111, SEQ ID NO:113, SEQID NO:121, SEQ ID NO:123, SEQ ID NO:l25, SEQ ID NO:127, SEQ ID NO:129,SEQ ID NO:131, SEQ ID NO:133, SEQ ID NO:135, SEQ ID NO:137, SEQ IDNO:139, SEQ ID NO:141, SEQ ID NO:143, SEQ ID NO:145, SEQ ID NO:147, SEQID NO:149, SEQ ID NO:151, SEQ ID NO:153, SEQ ID NO:155, SEQ ID NO:157,SEQ ID NO:159, SEQ ID NO:161, SEQ ID NO:163, SEQ ID NO:165, SEQ IDNO:179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:184 and/or SEQ IDNO:190. Additional particularly preferred serine protease proteins areencoded by allelic variants of nucleic acid molecules encoding proteinsthat include the cited amino acid sequences Also preferred are fleaserine protease proteins including regions that have at least about 50%,preferably at least about 75%, and more preferably at least about 90%identity with flea serine protease proteins having a amino acidsequences as cited herein.

One embodiment of the present invention is a flea serine protease thatdegrades IgG antibodies circulating in a host animal (i.e., fleaIgGase). An example of a flea IgGase is presented in the Examplessection.

A preferred flea aminopeptidase proteins of the present invention isencoded by a nucleic acid molecule that hybridizes under stringenthybridization conditions with nfAP and/or nfAP2. Particularly preferredflea aminopeptidase proteins of the present invention are encoded by anucleic acid molecule that hybridizes under stringent hybridizationconditions with nucleic acid molecule nfAP₄₅₃, nfAP₁₅₈₀ and/or nfAP2₅₃₇,the production of which are described in the Examples. Even morepreferred is an aminopeptidase that includes amino acid sequence SEQ IDNO:51, SEQ ID NO:113, SEQ ID NO:167, SEQ ID NO:170 and/or SEQ ID NO:172or an aminopeptidase encoded by an allelic variant of a nucleic acidmolecule that includes SEQ ID NO:50, SEQ ID NO:112, SEQ ID NO:169 and/orSEQ ID NO:171. Also preferred are flea aminopeptidase proteins includingregions that have at least about 50%, preferably at least about 75%, andmore preferably at least about 90% identity with SEQ ID NO:51, SEQ IDNO:113, SEQ ID NO:167, SEQ ID NO:170 and/or SEQ ID NO:172.

A preferred flea cysteine protease protein of the present invention isencoded by a nucleic acid molecule that hybridizes under stringenthybridization conditions with nucleic acid molecule nfCP1 (a fleacysteine protease full-length coding region that includes nfCP1₅₇₃, theproduction of which is described in the Examples). Even more preferredis a cysteine protease that includes amino acid sequence SEQ ID NO:178or a cysteine protease encoded by an allelic variant of a nucleic acidmolecule that includes SEQ ID NO:177. Also preferred is a flea cysteineprotease protein including regions that have at least about 50%,preferably at least about 75%, and more preferably at least about 90%identity with SEQ ID NO:178.

One embodiment of the present invention is an isolated protein havingproteolytic activity that is substantially inhibited by a serineprotease inhibitor, an aminopeptidase inhibitor and/or a cysteineprotease inhibitor. Such inhibition can be measured by techniques knownto those skilled in the art. To be substantially inhibited means, forexample, for a serine protease, that at least half of the proteolyticactivity of the protease protein is inhibited by a serine proteaseinhibitor. Preferably at least about 70 percent, and even morepreferably at least about 90 percent of the proteolytic activity of theprotease protein is inhibited by a serine protease inhibitor.

An isolated protein of the present invention can be produced in avariety of ways, including recovering such a protein from a flea midgutand producing such a protein recombinantly. In one embodiment, a fleamidgut protease can be recovered by methods heretofore disclosed forobtaining a soluble flea midgut preparation. A flea midgut proteaseprotein can be further purified from a disrupted flea midgut by a numberof techniques known to those skilled in the art, including, but notlimited to, affinity chromatography, ion exchange chromatography,filtration, electrophoresis (e.g., standard, capillary and flow-throughelectrophoresis), hydrophobic interaction chromatography, gel filtrationchromatography, reverse phase chromatography, concanavalin Achromatography, chromatofocusing and differential solubilization. In oneembodiment, a flea midgut protease is purified using protease inhibitoraffinity chromatography, an example of which is disclosed in theExamples section.

Another embodiment of the present invention is a method to produce anisolated protein of the present invention using recombinant DNAtechnology. Such a method includes the steps of (a) culturing arecombinant cell comprising a nucleic acid molecule encoding a proteinof the present invention to produce the protein and (b) recovering theprotein therefrom. Details on producing recombinant cells and culturingthereof are presented below. The phrase "recovering the protein" referssimply to collecting the whole fermentation medium containing theprotein and need not imply additional steps of separation orpurification. Proteins of the present invention can be purified using avariety of standard protein purification techniques, as heretoforedisclosed.

Isolated proteins of the present invention are preferably retrieved in"substantially pure" form. As used herein, "substantially pure" refersto a purity that allows for the effective use of the protein as avaccine. A vaccine for animals, for example, should exhibit nosubstantial toxicity and should be capable of stimulating the productionof antibodies in a vaccinated animal.

Another embodiment of the present invention is an isolated nucleic acidmolecule capable of hybridizing under stringent conditions with a geneencoding a flea protease present in a flea midgut. Such a nucleic acidmolecule is also referred to herein as a flea protease nucleic acidmolecule. Particularly preferred is an isolated nucleic acid moleculethat hybridizes under stringent conditions with a flea serine proteasegene, with a flea aminopeptidase gene or with a flea cysteine proteasegene. The characteristics of such genes are disclosed herein. Inaccordance with the present invention, an isolated nucleic acid moleculeis a nucleic acid molecule that has been removed from its natural milieu(i.e., that has been subject to human manipulation). As such, "isolated"does not reflect the extent to which the nucleic acid molecule has beenpurified. An isolated nucleic acid molecule can include DNA, RNA, orderivatives of either DNA or RNA.

As stated above, a flea protease gene includes all nucleic acidsequences related to a natural flea protease gene such as regulatoryregions that control production of a flea protease protein encoded bythat gene (such as, but not limited to, transcription, translation orpost-translation control regions) as well as the coding region itself. Anucleic acid molecule of the present invention can be an isolatednatural flea protease nucleic acid molecule or a homologue thereof. Anucleic acid molecule of the present invention can include one or moreregulatory regions, full-length or partial coding regions, orcombinations thereof. The minimal size of a flea protease nucleic acidmolecule of the present invention is the minimal size capable of forminga stable hybrid under stringent hybridization conditions with acorresponding natural gene. Flea protease nucleic acid molecules canalso include a nucleic acid molecule encoding a hybrid protein, a fusionprotein, a multivalent protein or a truncation fragment.

An isolated nucleic acid molecule of the present invention can beobtained from its natural source either as an entire (i.e., complete)gene or a portion thereof capable of forming a stable hybrid with thatgene. As used herein, the phrase "at least a portion of" an entityrefers to an amount of the entity that is at least sufficient to havethe functional aspects of that entity. For example, at least a portionof a nucleic acid sequence, as used herein, is an amount of a nucleicacid sequence capable of forming a stable hybrid with the correspondinggene under stringent hybridization conditions.

An isolated nucleic acid molecule of the present invention can also beproduced using recombinant DNA technology (e.g., polymerase chainreaction (PCR) amplification, cloning) or chemical synthesis. Isolatedflea protease nucleic acid molecules include natural nucleic acidmolecules and homologues thereof, including, but not limited to, naturalallelic variants and modified nucleic acid molecules in whichnucleotides have been inserted, deleted, substituted, and/or inverted insuch a manner that such modifications do not substantially interferewith the nucleic acid molecule's ability to encode a flea proteaseprotein of the present invention or to form stable hybrids understringent conditions with natural nucleic acid molecule isolates.

A flea protease nucleic acid molecule homologue can be produced using anumber of methods known to those skilled in the art (see, for example,Sambrook et al., ibid.). For example, nucleic acid molecules can bemodified using a variety of techniques including, but not limited to,classic mutagenesis techniques and recombinant DNA techniques, such assite-directed mutagenesis, chemical treatment of a nucleic acid moleculeto induce mutations, restriction enzyme cleavage of a nucleic acidfragment, ligation of nucleic acid fragments, polymerase chain reaction(PCR) amplification and/or mutagenesis of selected regions of a nucleicacid sequence, synthesis of oligonucleotide mixtures and ligation ofmixture groups to "build" a mixture of nucleic acid molecules andcombinations thereof. Nucleic acid molecule homologues can be selectedfrom a mixture of modified nucleic acids by screening for the functionof the protein encoded by the nucleic acid (e.g., the ability of ahomologue to elicit an immune response against a flea protease and/or tohave proteolytic activity) and/or by hybridization with isolated fleaprotease nucleic acids under stringent conditions.

An isolated flea protease nucleic acid molecule of the present inventioncan include a nucleic acid sequence that encodes at least one fleaprotease protein of the present invention, examples of such proteinsbeing disclosed herein. Although the phrase "nucleic acid molecule"primarily refers to the physical nucleic acid molecule and the phrase"nucleic acid sequence" primarily refers to the sequence of nucleotideson the nucleic acid molecule, the two phrases can be usedinterchangeably, especially with respect to a nucleic acid molecule, ora nucleic acid sequence, being capable of encoding an flea proteaseprotein.

One embodiment of the present invention is a flea protease nucleic acidmolecule of the present invention that is capable of hybridizing understringent conditions to a nucleic acid strand that encodes at least aportion of a flea protease or a homologue thereof or to the complementof such a nucleic acid strand. A nucleic acid sequence complement of anynucleic acid sequence of the present invention refers to the nucleicacid sequence of the nucleic acid strand that is complementary to (i.e.,can form a complete double helix with) the strand for which the sequenceis cited. It is to be noted that a double-stranded nucleic acid moleculeof the present invention for which a nucleic acid sequence has beendetermined for one strand, that is represented by a SEQ ID NO, alsocomprises a complementary strand having a sequence that is a complementof that SEQ ID NO. As such, nucleic acid molecules of the presentinvention, which can be either double-stranded or single-stranded,include those nucleic acid molecules that form stable hybrids understringent hybridization conditions with either a given SEQ ID NO denotedherein and/or with the complement of that SEQ ID NO, which may or maynot be denoted herein. Methods to deduce a complementary sequence areknown to those skilled in the art. Preferred is a flea protease nucleicacid molecule that includes a nucleic acid sequence having at leastabout 65 percent, preferably at least about 75 percent, more preferablyat least about 85 percent, and even more preferably at least about 95percent homology with the corresponding region(s) of the nucleic acidsequence encoding at least a portion of a flea protease protein.Particularly preferred is a flea protease nucleic acid molecule capableof encoding at least a portion of a flea protease that naturally ispresent in flea midguts and preferably is included in a soluble fleamidgut preparation of the present invention. Examples of nucleic acidmolecules of the present invention are disclosed in the Examplessection.

A preferred flea serine protease nucleic acid molecule of the presentinvention is a nucleic acid molecule that hybridizes under stringenthybridization conditions with at least one of the following nucleic acidmolecules: nfSP1, nfSP2, nfSP3, nfSP4, nfSP5, nfSP6, nfSP7, nfSP8,nfSP9, nfSP10, nfSP11, nfSP12, nfSP13, nfSP14, nfSP15, nfSP16, nfSP17,nfSP18, nfSP19, nfSP20, nfSP21, nfSP23, nfSP24, nfSP25, nfSP26, nfSP27,nfSP28, nfSP29, nfSP30, nfSP31, nfSP32, nfSP33, nfSP34, nfSP36, nfSP37,nfSP38 and/or nfSP39. More preferred is a nucleic acid molecule thathybridizes under stringent hybridization conditions with at least one ofthe following nucleic acid molecules: nfSP4₆₇₂, nfSP1₁₅₆, nfSP2₁₆₈,nfSP3₁₇₇, nfSP4₁₅₆, nfSP5₁₅₉, nfSP6₁₆₈, nfSP7₁₅₉, nfSP8₁₈₆, nfSP9₁₆₈,nfSP10₁₂₀, nfSP11₁₆₂, nfSP5₈₀₆, nfSP11₃₀₇, nfSP8₅₁₅, nfSP12₇₅₈,nfSP26₆₁₀, nfSP27₃₈₆, nfSP23₄₂₃, nfSP24₄₁₀, nfSP33₄₂₆, nfSP34₃₉₀,nfSP36₁₉₇, nfSP38₃₄₁, nfSP37₂₆₁, nfSP39₂₆₇ nfSP29₆₁₂, nfSP30₆₄₁,nfSP31₆₂₆, nfSP32₄₃₃, nfSP15₈₁₅, nfSP19₈₅₅, nfSP25₈₆₄, nfSP21₅₉₅,nfSP28₉₂₃ and/or nfSP33₇₇₈, as well as other specific nucleic acidmolecules disclosed in the Examples section. Even more preferred arenucleic acid molecules that include nfSP1, nfSP2, nfSP3, nfSP4, nfSP5,nfSP6, nfSP7, nfSP8, nfSP9, nfSP10, nfSP11, nfSP12, nfSP13, nfSP14,nfSP15, nfSP16, nfSP17, nfSP18, nfSP19, nfSP20, nfSP21, nfSP23, nfSP24,nfSP25, nfSP26, nfSP27, nfSP28, nfSP29, nfSP30, nfSP31, nfSP32, nfSP33,nfSP34, nfSP36, nfSP37, nfSP38 and/or nfSP39 and even more particularly,nfSP4₆₇₂, nfSP1₁₅₆, nfSP2₁₆₈, nfSP3₁₇₇, nfSP4₁₅₆, nfSP5₁₅₉, nfSP6₁₆₈,nfSP7₁₅₉, nfSP8₁₈₆, nfSP9₁₆₈, nfSP10₁₂₀₁, nfSP11₁₆₂, nfSP5₈₀₆,nfSP11₃₀₇, nfSP8₅₁₅, nfSP12₇₅₈, nfSP26₆₁₀, nfSP27₃₈₆, nfSP23₄₂₃,nfSP24₄₁₀, nfSP33₄₂₆, nfSP34₃₉₀, nfSP36₁₉₇, nfSP38₃₄₁, nfSP37₂₆₁,nfSP39₂₆₇ nfSP29₆₁₂, nfSP30₆₄₁, nfSP31₆₂₆, nfSP32₄₃₃, nfSP15₈₁₅,nfSP19₈₅₅, nfSP25₈₆₄, nfSP21₅₉₅, nfSP28₉₂₃, nfSP33₇₇₈, as well as otherspecific nucleic acid molecules disclosed in the Examples section.

Particularly preferred flea serine protease nucleic acid moleculesinclude at least one of the following sequences: SEQ ID NO:16, SEQ IDNO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ IDNO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ IDNO:38, and/or nucleic acid sequences disclosed in Table 2 (i.e., SEQ IDNO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ IDNO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ IDNO:72, SEQ ID NO:74, SEQ ID NO:76, SEQ ID NO:78), as well as SEQ IDNO:80, SEQ ID NO:82, SEQ ID NO:84, SEQ ID NO:86, SEQ ID NO:88, SEQ IDNO:90, SEQ ID NO:92, SEQ ID NO:94, SEQ ID NO:96, SEQ ID NO:98, SEQ IDNO:100, SEQ ID NO:102, SEQ ID NO:104, SEQ ID NO:106, SEQ ID NO:108, SEQID NO:110, SEQ ID NO:120, SEQ ID NO:122, SEQ ID NO:124, SEQ ID NO:126,SEQ ID NO:128, SEQ ID NO:130, SEQ ID NO:132, SEQ ID NO:134, SEQ IDNO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, SEQID NO:146, SEQ ID NO:148, SEQ ID NO:150, SEQ ID NO:152, SEQ ID NO:154,SEQ ID NO:156, SEQ ID NO:158, SEQ ID NO:160, SEQ ID NO:162, SEQ IDNO:164 and/or SEQ ID NO:189, and complements thereof. Also preferred areallelic variants of such nucleic acid molecules.

A preferred flea aminopeptidase nucleic acid molecule of the presentinvention is a nucleic acid molecule that hybridizes under stringenthybridization conditions with nfAP and/or nfAP2. A more preferred fleaaminopeptidase nucleic acid molecule of the present invention is anucleic acid molecule that hybridizes under stringent hybridizationconditions with nfAP₄₅₃, nfAP₉₀₀, nfAP₇₃₂, nfAP₁₅₈₀, nfAP2₃₈₃ and/ornfAP2₅₃₇. More preferred is an aminopeptidase nucleic acid molecule thatincludes nfAP₄₅₃, nfAP₉₀₀, nfAP₇₃₂, nfAP₁₅₈₀, nfAP2₃₈₃ and/or nfAP2₅₃₇.Particularly preferred is a nucleic acid molecule that includes nucleicacid sequence SEQ ID NO:50, SEQ ID NO:112, SEQ ID NO:169 and/or SEQ IDNO:171, a complement of any of such sequences, or allelic variantsthereof.

A preferred flea cysteine protease nucleic acid molecule of the presentinvention is a nucleic acid molecule that hybridizes under stringenthybridization conditions with nfCP1₅₇₃. More preferred is a cysteineprotease nucleic acid molecule that includes nfCP1₅₇₃. Particularlypreferred is a nucleic acid molecule that includes nucleic acid sequenceSEQ ID NO:177, a complement of SEQ ID NO:177, or allelic variants ofsuch nucleic acid molecule.

Knowing a nucleic acid molecule of a flea protease protein of thepresent invention allows one skilled in the art to make copies of thatnucleic acid molecule as well as to obtain a nucleic acid moleculeincluding additional portions of flea protease protein-encoding genes(e.g., nucleic acid molecules that include the translation start siteand/or transcription and/or translation control regions), and/or fleaprotease nucleic acid molecule homologues. Knowing a portion of an aminoacid sequence of a flea protease protein of the present invention allowsone skilled in the art to clone nucleic acid sequences encoding such aflea protease protein. In addition, a desired flea protease nucleic acidmolecule can be obtained in a variety of ways including screeningappropriate expression libraries with antibodies which bind to fleaprotease proteins of the present invention; traditional cloningtechniques using oligonucleotide probes of the present invention toscreen appropriate libraries or DNA; and PCR amplification ofappropriate libraries, or RNA or DNA using oligonucleotide primers ofthe present invention (genomic and/or cDNA libraries can be used). Toisolate flea protease nucleic acid molecules, preferred cDNA librariesinclude cDNA libraries made from unfed whole fleas, fed whole fleas, fedflea midguts, unfed flea midguts, and flea salivary glands. Techniquesto clone and amplify genes are disclosed, for example, in Sambrook etal., ibid. The Examples section includes examples of the isolation ofcDNA sequences encoding flea protease proteins of the present invention.

The present invention also includes nucleic acid molecules that areoligonucleotides capable of hybridizing, under stringent conditions,with complementary regions of other, preferably longer, nucleic acidmolecules of the present invention that encode at least a portion of aflea protease protein. Oligonucleotides of the present invention can beRNA, DNA, or derivatives of either. The minimal size of sucholigonucleotides is the size required to form a stable hybrid between agiven oligonucleotide and the complementary sequence on another nucleicacid molecule of the present invention. Minimal size characteristics aredisclosed herein. The size of the oligonucleotide must also besufficient for the use of the oligonucleotide in accordance with thepresent invention. Oligonucleotides of the present invention can be usedin a variety of applications including, but not limited to, as probes toidentify additional nucleic acid molecules, as primers to amplify orextend nucleic acid molecules or in therapeutic applications to inhibitflea protease production. Such therapeutic applications include the useof such oligonucleotides in, for example, antisense-, triplexformation-, ribozyme- and/or RNA drug-based technologies. The presentinvention, therefore, includes such oligonucleotides and methods tointerfere with the production of flea protease proteins by use of one ormore of such technologies.

The present invention also includes a recombinant vector, which includesa flea protease nucleic acid molecule of the present invention insertedinto any vector capable of delivering the nucleic acid molecule into ahost cell. Such a vector contains heterologous nucleic acid sequences,that is nucleic acid sequences that are not naturally found adjacent toflea protease nucleic acid molecules of the present invention. Thevector can be either RNA or DNA, either prokaryotic or eukaryotic, andtypically is a virus or a plasmid. Recombinant vectors can be used inthe cloning, sequencing, and/or otherwise manipulating of flea proteasenucleic acid molecules of the present invention. One type of recombinantvector, herein referred to as a recombinant molecule and described inmore detail below, can be used in the expression of nucleic acidmolecules of the present invention. Preferred recombinant vectors arecapable of replicating in the transformed cell. Preferred nucleic acidmolecules to include in recombinant vectors of the present invention aredisclosed herein.

As heretofore disclosed, one embodiment of the present invention is amethod to produce a flea protease protein of the present invention byculturing a cell capable of expressing the protein under conditionseffective to produce the protein, and recovering the protein. Apreferred cell to culture is a recombinant cell that is capable ofexpressing the flea protease protein, the recombinant cell beingproduced by transforming a host cell with one or more nucleic acidmolecules of the present invention. Transformation of a nucleic acidmolecule into a cell can be accomplished by any method by which anucleic acid molecule can be inserted into the cell. Transformationtechniques include, but are not limited to, transfection,electroporation, microinjection, lipofection, adsorption, and protoplastfusion. A recombinant cell may remain unicellular or may grow into atissue, organ or a multicellular organism. Transformed nucleic acidmolecules of the present invention can remain extrachromosomal or canintegrate into one or more sites within a chromosome of the transformed(i.e., recombinant) cell in such a manner that their ability to beexpressed is retained. Preferred nucleic acid molecules with which totransform a host cell are disclosed herein.

Suitable host cells to transform include any cell that can betransformed and that can express the introduced flea protease protein.Such cells are, therefore, capable of producing flea protease proteinsof the present invention after being transformed with at least onenucleic acid molecule of the present invention. Host cells can be eitheruntransformed cells or cells that are already transformed with at leastone nucleic acid molecule. Suitable host cells of the present inventioncan include bacterial, fungal (including yeast), insect, animal andplant cells. Preferred host cells include bacterial, yeast, insect andmammalian cells, with bacterial (e.g., E. coli) and insect (e.g.,Spodoptera) cells being particularly preferred.

A recombinant cell is preferably produced by transforming a host cellwith one or more recombinant molecules, each comprising one or morenucleic acid molecules of the present invention operatively linked to anexpression vector containing one or more transcription controlsequences. The phrase operatively linked refers to insertion of anucleic acid molecule into an expression vector in a manner such thatthe molecule is able to be expressed when transformed into a host cell.As used herein, an expression vector is a DNA or RNA vector that iscapable of transforming a host cell and of effecting expression of aspecified nucleic acid molecule. Preferably, the expression vector isalso capable of replicating within the host cell. Expression vectors canbe either prokaryotic or eukaryotic, and are typically viruses orplasmids. Expression vectors of the present invention include anyvectors that function (i.e., direct gene expression) in recombinantcells of the present invention, including in bacterial, fungal, insect,animal, and/or plant cells. As such, nucleic acid molecules of thepresent invention can be operatively linked to expression vectorscontaining regulatory sequences such as promoters, operators,repressors, enhancers, termination sequences, origins of replication,and other regulatory sequences that are compatible with the recombinantcell and that control the expression of nucleic acid molecules of thepresent invention. As used herein, a transcription control sequenceincludes a sequence which is capable of controlling the initiation,elongation, and termination of transcription. Particularly importanttranscription control sequences are those which control transcriptioninitiation, such as promoter, enhancer, operator and repressorsequences. Suitable transcription control sequences include anytranscription control sequence that can function in at least one of therecombinant cells of the present invention. A variety of suchtranscription control sequences are known to those skilled in the art.Preferred transcription control sequences include those which functionin bacterial, yeast, helminth, insect and mammalian cells, such as, butnot limited to, tac, lac, trp, trc, oxy-pro, omp/lpp, rrnB,bacteriophage lambda (λ) (such as λP_(L) and λP_(R) and fusions thatinclude such promoters), bacteriophage T7, T7lac, bacteriophage T3,bacteriophage SP6, bacteriophage SP01, metallothionein, alpha matingfactor, Pichia alcohol oxidase, alphavirus subgenomic promoters (such asSindbis virus subgenomic promoters), baculovirus, Heliothis zea insectvirus, vaccinia virus, herpesvirus, poxvirus, adenovirus, simian virus40, retrovirus actin, retroviral long terminal repeat, Rous sarcomavirus, heat shock, phosphate and nitrate transcription control sequencesas well as other sequences capable of controlling gene expression inprokaryotic or eukaryotic cells. Additional suitable transcriptioncontrol sequences include tissue-specific promoters and enhancers aswell as lymphokine-inducible promoters (e.g., promoters inducible byinterferons or interleukins). Transcription control sequences of thepresent invention can also include naturally occurring transcriptioncontrol sequences naturally associated with a DNA sequence encoding aflea protease protein.

Expression vectors of the present invention may also contain secretorysignals (i.e., signal segment nucleic acid sequences) to enable anexpressed flea protease protein to be secreted from the cell thatproduces the protein. Suitable signal segments include a flea proteaseprotein signal segment or any heterologous signal segment capable ofdirecting the secretion of a flea protease protein, including fusionproteins, of the present invention. Preferred signal segments include,but are not limited to, flea protease, tissue plasminogen activator(t-PA), interferon, interleukin, growth hormone, histocompatibility andviral envelope glycoprotein signal segments.

Expression vectors of the present invention may also contain fusionsequences which lead to the expression of inserted nucleic acidmolecules of the present invention as fusion proteins. Inclusion of afusion sequence as part of a flea protease nucleic acid molecule of thepresent invention can enhance the stability during production, storageand/or use of the protein encoded by the nucleic acid molecule.Furthermore, a fusion segment can function as a tool to simplifypurification of a flea protease protein, such as to enable purificationof the resultant fusion protein using affinity chromatography. Asuitable fusion segment can be a domain of any size that has the desiredfunction (e.g., increased stability and/or purification tool). It iswithin the scope of the present invention to use one or more fusionsegments. Fusion segments can be joined to amino and/or carboxyl terminiof a flea protease protein. Linkages between fusion segments and fleaprotease proteins can be constructed to be susceptible to cleavage toenable straight-forward recovery of the flea protease proteins. Fusionproteins are preferably produced by culturing a recombinant celltransformed with a fusion nucleic acid sequence that encodes a proteinincluding the fusion segment attached to either the carboxyl and/oramino terminal end of a flea protease protein.

A recombinant molecule of the present invention is a molecule that caninclude at least one of any nucleic acid molecule heretofore describedoperatively linked to at least one of any transcription control sequencecapable of effectively regulating expression of the nucleic acidmolecule(s) in the cell to be transformed. A preferred recombinantmolecule includes one or more nucleic acid molecules of the presentinvention, with those that encode one or more flea protease proteins,and particularly one or more flea serine protease, aminopeptidase and/orcysteine protease proteins, being more preferred. Similarly, a preferredrecombinant cell includes one or more nucleic acid molecules of thepresent invention, with those that encode one or more flea proteaseproteins, and particularly one or more flea serine protease,aminopeptidase, and/or cysteine protease proteins, being more preferred.

It may be appreciated by one skilled in the art that use of recombinantDNA technologies can improve expression of transformed nucleic acidmolecules by manipulating, for example, the number of copies of thenucleic acid molecules within a host cell, the efficiency with whichthose nucleic acid molecules are transcribed, the efficiency with whichthe resultant transcripts are translated, and the efficiency ofpost-translational modifications. Recombinant techniques useful forincreasing the expression of nucleic acid molecules of the presentinvention include, but are not limited to, operatively linking nucleicacid molecules to high-copy number plasmids, integration of the nucleicacid molecules into one or more host cell chromosomes, addition ofvector stability sequences to plasmids, substitutions or modificationsof transcription control signals (e.g., promoters, operators,enhancers), substitutions or modifications of translational controlsignals (e.g., ribosome binding sites, Shine-Dalgarno sequences),modification of nucleic acid molecules of the present invention tocorrespond to the codon usage of the host cell, deletion of sequencesthat destabilize transcripts, and use of control signals that temporallyseparate recombinant cell growth from recombinant protein productionduring fermentation. The activity of an expressed recombinant protein ofthe present invention may be improved by fragmenting, modifying, orderivatizing the resultant protein.

In accordance with the present invention, recombinant cells can be usedto produce flea protease proteins of the present invention by culturingsuch cells under conditions effective to produce such a protein, andrecovering the protein. Effective conditions to produce a proteininclude, but are not limited to, appropriate media, bioreactor,temperature, pH and oxygen conditions that permit protein production. Anappropriate, or effective, medium refers to any medium in which a cellof the present invention, when cultured, is capable of producing a fleaprotease protein. Such a medium is typically an aqueous mediumcomprising assimilable carbohydrate, nitrogen and phosphate sources, aswell as appropriate salts, minerals, metals and other nutrients, such asvitamins. The medium may comprise complex nutrients or may be a definedminimal medium.

Cells of the present invention can be cultured in conventionalfermentation bioreactors, which include, but are not limited to, batch,fed-batch, cell recycle, and continuous fermentors. Culturing can alsobe conducted in shake flasks, test tubes, microtiter dishes, and petriplates. Culturing is carried out at a temperature, pH and oxygen contentappropriate for the recombinant cell. Such culturing conditions are wellwithin the expertise of one of ordinary skill in the art.

Depending on the vector and host system used for production, resultantflea protease proteins may either remain within the recombinant cell; besecreted into the fermentation medium; be secreted into a space betweentwo cellular membranes, such as the periplasmic space in E. coli; or beretained on the outer surface of a cell or viral membrane. Methods topurify such proteins are heretofore disclosed.

The present invention also includes isolated anti-flea proteaseantibodies and their use to reduce flea infestation on a host animal aswell as in the environment of the animal. An anti-flea protease antibodyis an antibody capable of selectively binding to a protease present in aflea midgut, including female and male fed midguts as well as female andmale unfed midguts. An anti-flea protease antibody preferably binds tothe protease in such a way as to reduce the proteolytic activity of thatprotease.

Isolated antibodies are antibodies that have been removed from theirnatural milieu. The term "isolated" does not refer to the state ofpurity of such antibodies. As such, isolated antibodies can includeanti-sera containing such antibodies, or antibodies that have beenpurified to varying degrees. As used herein, the term "selectively bindsto" refers to the ability of such antibodies to preferentially bind tothe protease against which the antibody was raised (i.e., to be able todistinguish that protease from unrelated components in a mixture.).Binding affinities typically range from about 10³ M⁻¹ to about 10¹² M⁻¹.Binding can be measured using a variety of methods known to thoseskilled in the art including immunoblot assays, immunoprecipitationassays, radioimmunoassays, enzyme immunoassays (e.g., ELISA),immunofluorescent antibody assays and immunoelectron microscopy; see,for example, Sambrook et al., ibid.

Antibodies of the present invention can be either polyclonal ormonoclonal antibodies. Antibodies of the present invention includefunctional equivalents such as antibody fragments andgenetically-engineered antibodies, including single chain antibodies,that are capable of selectively binding to at least one of the epitopesof the protein used to obtain the antibodies. Antibodies of the presentinvention also include chimeric antibodies that can bind to more thanone epitope. Preferred antibodies are raised in response to proteinsthat are encoded, at least in part, by a flea protease nucleic acidmolecule of the present invention.

Anti-flea antibodies of the present invention include antibodies raisedin an animal administered a flea protease vaccine of the presentinvention that exert their effect when fleas feed from the vaccinatedanimal's blood containing such antibodies. Anti-flea antibodies of thepresent invention also include antibodies raised in an animal againstone or more flea protease proteins, or soluble flea midgut preparations,of the present invention that are then recovered from the animal usingtechniques known to those skilled in the art. Yet additional antibodiesof the present invention are produced recombinantly using techniques asheretofore disclosed for flea protease proteins of the presentinvention. Antibodies produced against defined proteins can beadvantageous because such antibodies are not substantially contaminatedwith antibodies against other substances that might otherwise causeinterference in a diagnostic assay or side effects if used in atherapeutic composition.

Anti-flea protease antibodies of the present invention have a variety ofuses that are within the scope of the present invention. For example,such antibodies can be used in a composition of the present invention topassively immunize an animal in order to protect the animal from fleainfestation. Anti-flea antibodies can also be used as tools to screenexpression libraries and/or to recover desired proteins of the presentinvention from a mixture of proteins and other contaminants.Furthermore, antibodies of the present invention can be used to targetcytotoxic agents to fleas in order to kill fleas. Targeting can beaccomplished by conjugating (i.e., stably joining) such antibodies tothe cytotoxic agents using techniques known to those skilled in the art.

A preferred anti-flea protease antibody of the present invention canselectively bind to, and preferentially reduce the proteolytic activityof, a flea serine protease, a flea metalloprotease, a flea aspartic acidprotease and/or a flea cysteine protease. More preferred anti-fleaprotease antibodies include anti-flea serine protease antibodies,anti-flea metalloprotease antibodies, anti-flea aminopeptidaseantibodies, and anti-flea cysteine protease antibodies. Particularlypreferred are anti-flea serine protease antibodies, anti-fleaaminopeptidase antibodies, and anti-flea cysteine protease antibodies,including those raised against flea serine protease proteins, fleaaminopeptidase proteins or cysteine protease proteins of the presentinvention.

The present invention also includes the use of protease inhibitors thatreduce proteolytic activity of flea proteases to reduce flea infestationof animals and the surrounding environment. As used herein, proteaseinhibitors are compounds that interact directly with a protease therebyinhibiting that protease's activity, usually by binding to or otherwiseinteracting with the protease's active site. Protease inhibitors areusually relatively small compounds and as such differ from anti-proteaseantibodies that interact with the active site of a protease.

Protease inhibitors can be used directly as compounds in compositions ofthe present invention to treat animals as long as such compounds are notharmful to the animals being treated. Protease inhibitors can also beused to identify preferred types of flea proteases to target usingcompositions of the present invention. For example, the inventors haveshown herein the predominance of serine proteases in flea midguts,particularly in soluble flea midgut preparations, using proteaseinhibitors. Such knowledge suggests that effective reduction of fleainfestation of an animal can be achieved using serine protease vaccines,anti-flea serine protease antibodies and other inhibitors of serineprotease synthesis and activity that can be tolerated by the animal. Forexample, flea IgGase activity disclosed herein can be targeted to reduceflea infestation. That other proteases are also present in flea midgutsaccording to the present invention also suggests targeting suchproteases. Methods to use protease inhibitors are known to those skilledin the art; examples of such methods are disclosed herein.

In one embodiment, a protease inhibitor that can be used in acomposition of the present invention to treat an animal is identified bya method including the following steps: (a) identifying candidate (i.e.,putative, possible) inhibitor compounds by testing the efficacy of oneor more protease inhibitors (i) in vitro for their ability to inhibitflea protease activity and/or (ii) in a flea feeding assay for theirability to reduce the survival and/or fecundity of fleas by adding theinhibitors to the blood meal of a flea being maintained, for example, ina feeding system, such as that described by Wade et al., 1988, J.MedEntomol. 25, 186-190; and (b) testing the efficacy of the candidateinhibitor compounds in animals infested with fleas. Although one doesnot need both in vitro assay data and flea feeding assay data todetermine which candidate compounds to administer to animals, evaluationof both sets of data is preferred since data from neither of the assaysnecessarily predicts data to be obtained from the other assay. Forexample, candidate compounds identified using the in vitro assay maywork "in the test tube" but may not work in vivo for a number ofreasons, including the presence of interfering components in the bloodmeal that inhibit the activity of such compounds; e.g., althoughaprotinin can inhibit at least some flea serine proteases in vitro,aprotinin does not work well in the presence of serum proteins, such asare found in the blood. Furthermore, candidate inhibitor compoundsidentified by the flea feeding assays can include not only desiredcompounds but also compounds that reduce the viability and/or fecundityof fleas due to general toxicity (e.g., affecting the mitochondria offleas).

In another embodiment, protease inhibitors are used in the purificationof corresponding proteases by, for example, affinity chromatography, inwhich, a protease inhibitor is incubated with a mixture containing adesired protease under conditions that the inhibitor forms a complexwith the protease. The protease can then be recovered from the complex.The protease inhibitor can be attached to a solid support and/or belabelled with, for example, a radioactive, fluorescent, or enzymatic tagthat can be used to detect and/or recover the complex.

Suitable protease inhibitors to use in accordance with the presentinvention include serine protease inhibitors (including IgGaseinhibitors) , metalloprotease inhibitors, aspartic acid proteaseinhibitors, cysteine protease inhibitors and aminopeptidase inhibitors.Preferred protease inhibitors include serine protease inhibitors,metalloprotease inhibitors, aminopeptidase inhibitors and cysteineprotease inhibitors, particularly those that are broad spectruminhibitors. More preferred are broad spectrum serine proteaseinhibitors.

There is a wide variety of protease inhibitors, as is known to oneskilled in the art. Examples include, but are not limited to, AEBSF,aprotinin, bestatin, chloromethyl ketones TLCK (Nα-p-tosyl-L-lysinechloromethyl ketone) and TPCK (N-tosyl-L-phenylalanine chloromethylketone), chymostatin, cystatin, 3'4-dichloroisocoumarin,E-64(trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane), EDTA(ethylenediaminetetraacetic acid), leupeptin, methyl ketones having avariety of leaving groups, oxidized L-leucinethiol, pepstatin,1,10-orthophenanthroline, phosphoramidon, soybean trypsin/chymotrypsininhibitor and soybean trypsin inhibitor. Preferred protease inhibitorsfor use in the present invention include AEBSF, bestatin, E-64leupeptin, pepstatin, 1,10-orthophenanthroline, phosphoramidon, TLCK andTPCK, with AEBSF (a broad spectrum serine protease inhibitor), bestatin(an inhibitor of leucine aminopeptidase) and 1,10-orthophenanthroline (abroad spectrum metalloprotease inhibitor) being particularly preferred.

Protease inhibitors can be produced using methods known to those skilledin the art. Protein- or peptide-based protease inhibitors, such ascystatin or small peptides comprising a protease substrate, can beproduced recombinantly and modified as necessary.

The present invention also includes the use of proteolytically activeflea protease proteins of the present invention to identify additionalprotease inhibitors, and preferably protease inhibitor compounds thatcan be included in a composition of the present invention to beadministered to animals. A method to identify a flea protease inhibitorincludes the steps of (a) contacting (e.g., combining, mixing) anisolated flea protease protein with a putative (i.e., candidate)inhibitory compound under conditions in which, in the absence of thecompound, the protein has proteolytic activity, and (b) determining ifthe putative inhibitory compound inhibits the proteolytic activity ofthe protein. Putative inhibitory compounds to screen include organicmolecules, antibodies (including functional equivalents thereof) andsubstrate analogs. Methods to determine protease activity are known tothose skilled in the art, as heretofore disclosed. Particularlypreferred for use in identifying inhibitors are flea serine proteaseproteins, flea aminopeptidase proteins and flea cysteine proteaseproteins of the present invention.

The present invention also includes a test kit to identify a compoundcapable of inhibiting flea protease activity. Such a test kit includesan isolated flea protease protein having proteolytic activity and ameans for determining the extent of inhibition of proteolytic activityin the presence of (i.e., effected by) a putative inhibitory compound.

The present invention also includes inhibitors isolated by such amethod, and/or test kit, and their use to inhibit any flea protease thatis susceptible to such an inhibitor.

It is to be appreciated that the present invention also includesmimetopes of compounds of the present invention that can be used inaccordance with methods as disclosed for compounds of the presentinvention. As used herein, a mimetope of a proteinaceous compound of thepresent invention (e.g., a flea protease protein, an anti-flea proteaseantibody, a proteinaceous inhibitor of protease activity or synthesis)refers to any compound that is able to mimic the activity of thatproteinaceous compound, often because the mimetope has a structure thatmimics the proteinaceous compound. For example, a mimetope of a fleaprotease protein is a compound that has an activity similar to that ofan isolated flea protease protein of the present invention. Mimetopescan be, but are not limited to: peptides that have been modified todecrease their susceptibility to degradation; anti-idiotypic and/orcatalytic antibodies, or fragments thereof; non-proteinaceousimmunogenic portions of an isolated protein (e.g., carbohydratestructures); and synthetic or natural organic molecules, includingnucleic acids. Such mimetopes can be designed using computer-generatedstructures of proteins of the present invention. Mimetopes can also beobtained by generating random samples of molecules, such asoligonucleotides, peptides or other organic molecules, and screeningsuch samples by affinity chromatography techniques using thecorresponding binding partner.

The present invention includes therapeutic compositions, also referredto herein as compositions, that include a (i.e., at least one) compoundof the present invention. Preferred compounds to include in acomposition of the present invention include flea protease vaccines,anti-flea protease antibodies and/or protease inhibitors as disclosedherein. Such a therapeutic composition can protect an animal from fleainfestation by reducing flea protease activity, thereby reducing fleaburden on the animal and in the environment of the animal.

Particularly preferred therapeutic compositions of the present inventioninclude at least one of the following compounds: an isolated flea serineprotease protein or a mimetope thereof; an isolated flea serine proteasenucleic acid molecule that hybridizes under stringent hybridizationconditions with a flea serine protease gene; an isolated antibody thatselectively binds to a flea serine protease protein; an inhibitor offlea serine protease activity identified by its ability to inhibit fleaserine protease activity; an isolated flea aminopeptidase protein or amimetope thereof; an isolated flea aminopeptidase nucleic acid moleculethat hybridizes under stringent hybridization conditions with a fleaaminopeptidase gene; an isolated antibody that selectively binds to aflea aminopeptidase protein; an inhibitor of flea aminopeptidaseactivity identified by its ability to inhibit flea aminopeptidaseactivity; an isolated flea cysteine protease protein or a mimetopethereof; an isolated flea cysteine protease nucleic acid molecule thathybridizes under stringent hybridization conditions with a flea cysteineprotease gene; an isolated antibody that selectively binds to a fleacysteine protease protein; and an inhibitor of flea cysteine proteaseactivity identified by its ability to inhibit flea cysteine proteaseactivity.

Another embodiment of the present invention is a therapeutic compositionthat includes a first compound that reduces flea protease activity and asecond compound that reduces flea burden by a method other than byreducing flea protease activity. The present invention also includes amethod to protect an animal from flea infestation by administering tothe animal such a composition. The first compound of such a compositionby effectively reducing flea protease activity in the midgut, enhancesthe activity of the second compound. While not being bound by theory, itis believed that a number of anti-flea treatments, particularly thosethat are proteinaceous, are not very effective because they are degradedin the flea midgut. The present invention permits the effective use ofsuch anti-flea treatments by reducing proteolytic degradation of suchtreatments by the flea midgut.

Preferred first compounds to include in such a composition include fleaprotease vaccines, anti-flea protease antibodies and/or proteaseinhibitors as disclosed herein, such compounds that target flea IgGaseactivity.

Suitable second compounds include any anti-flea agent(s), including, butnot limited to, proteinaceous compounds, insecticides and flea collars.Preferred second compounds are proteinaceous compounds that effectactive immunization (e.g., antigen vaccines), passive immunization(e.g., antibodies), or that otherwise inhibit a flea activity that wheninhibited can reduce flea burden on and around an animal. Examples ofsecond compounds include a compound that inhibits binding between a fleamembrane protein and its ligand (e.g., a compound that inhibits fleaATPase activity or a compound that inhibits binding of a peptide orsteroid hormone to its receptor), a compound that inhibits hormone(including peptide or steroid hormones) synthesis, a compound thatinhibits vitellogenesis (including production of vitellin and transportand maturation thereof into a major egg yolk protein), a compound thatinhibits fat body function, a compound that inhibits flea muscle action,a compound that inhibits the flea nervous system, a compound thatinhibits the flea immune system and/or a compound that inhibits fleafeeding.

Compositions of the present invention can also include other componentssuch as a pharmaceutically acceptable excipient, an adjuvant, and/or acarrier. For example, compositions of the present invention can beformulated in an excipient that the animal to be treated can tolerate.

Examples of such excipients include water, saline, Ringer's solution,dextrose solution, Hank's solution, and other aqueous physiologicallybalanced salt solutions. Nonaqueous vehicles, such as fixed oils, sesameoil, ethyl oleate, or triglycerides may also be used. Other usefulformulations include suspensions containing viscosity enhancing agents,such as sodium carboxymethylcellulose, sorbitol, or dextran. Excipientscan also contain minor amounts of additives, such as substances thatenhance isotonicity and chemical stability. Examples of buffers includephosphate buffer, bicarbonate buffer and Tris buffer, while examples ofpreservatives include thimerosal, m- or o-cresol, formalin and benzylalcohol. Standard formulations can either be liquid injectables orsolids which can be taken up in a suitable liquid as a suspension orsolution for injection. Thus, in a non-liquid formulation, the excipientcan comprise dextrose, human serum albumin, preservatives, etc., towhich sterile water or saline can be added prior to administration.

In one embodiment of the present invention, the composition can alsoinclude an immunopotentiator, such as an adjuvant or a carrier.Adjuvants are typically substances that generally enhance the immuneresponse of an animal to a specific antigen. Suitable adjuvants include,but are not limited to, Freund's adjuvant; other bacterial cell wallcomponents; aluminum-based salts; calcium-based salts; silica;polynucleotides; toxoids; serum proteins; viral coat proteins; otherbacterial-derived preparations; gamma interferon; block copolymeradjuvants, such as Hunter's Titermax adjuvant (Vaxcel™, Inc. Norcross,Ga.); Ribi adjuvants (available from Ribi ImmunoChem Research, Inc.,Hamilton, Mont.); and saponins and their derivatives, such as Quil A(available from Superfos Biosector A/S, Denmark). Carriers are typicallycompounds that increase the half-life of a therapeutic composition inthe treated animal. Suitable carriers include, but are not limited to,polymeric controlled release formulations, biodegradable implants,liposomes, bacteria, viruses, oils, esters, and glycols.

One embodiment of the present invention is a controlled releaseformulation that is capable of slowly releasing a composition of thepresent invention into an animal. As used herein a controlled releaseformulation comprises a composition of the present invention in acontrolled release vehicle. Suitable controlled release vehiclesinclude, but are not limited to, biocompatible polymers, other polymericmatrices, capsules, microcapsules, microparticles, bolus preparations,osmotic pumps, diffusion devices, liposomes, lipospheres, andtransdermal delivery systems. Other controlled release formulations ofthe present invention include liquids that, upon administration to ananimal, form a solid or a gel in situ. Preferred controlled releaseformulations are biodegradable (i.e., bioerodible).

A preferred controlled release formulation of the present invention iscapable of releasing a composition of the present invention into theblood of the treated animal at a constant rate sufficient to attaintherapeutic dose levels of the composition to reduce protease activityin fleas feeding from the animal over a period of time ranging fromabout 1 to about 12 months. A controlled release formulation of thepresent invention is capable of effecting a treatment for preferably atleast about 1 month, more preferably at least about 3 months and evenmore preferably for at least about 6 months, even more preferably for atleast about 9 months, and even more preferably for at least about 12months.

In order to protect an animal from flea infestation, a therapeuticcomposition of the present invention is administered to the animal in aneffective manner such that the protease activity of fleas feeding fromthe blood stream of animals treated with the composition is reduced. Assuch, a treated animal is an animal that is competent to reduce the fleaburden by reducing flea protease activity, or by reducing flea proteaseactivity and at least one other flea activity. Preferably, the proteaseactivity is reduced by at least about 50 percent, more preferably by atleast about 70 percent and even more preferably by at least about 90percent. Methods to administer compositions to the animal in order torender the animal competent depend on the nature of the composition andadministration regime. Animals administered a protease vaccine with atleast one booster shot usually become competent at about the same timeas would be expected for any vaccine treatment. For example, animalsadministered a booster dose about 4 to 6 weeks after a primary doseusually become competent within another about 3 to 4 weeks. Animalsadministered a composition including an anti-flea protease antibody orprotease inhibitor become competent as soon as appropriate serum levelsof the compound are achieved, usually with one to three days.

In a preferred embodiment, a composition of the present invention whenadministered to a host animal is able to reduce flea viability by atleast about 50 percent within at least about 21 days after the fleasbegin feeding from the treated animal. (Note that fleas usually liveabout 40 days to about 50 days on one or more animals.) A more preferredcomposition when administered to a host animal is able to reduce fleaviability by at least about 65 percent within at least about 14 daysafter the fleas begin feeding from the treated animal. An even morepreferred composition when administered to an animal is able to reduceflea viability by at least about 90 percent within at least about 7 daysafter the fleas begin feeding from the treated animal.

In another preferred embodiment, a composition of the present inventionwhen administered to a host animal is able to reduce flea fecundity(i.e., egg laying ability) by at least about 50 percent, more preferablyby at least about 70 percent, and even more preferably by at least about90 percent, within at least about 30 days after the fleas begin feedingfrom the treated animal. (Note that fleas usually do not begin layingeggs until about 7 days after taking a blood meal.)

In accordance with the present invention, compositions are administeredto an animal in a manner such that the animal becomes competent toreduce flea protease activity in a flea that feeds from the competent;i.e., the animal becomes a treated animal. For example, a flea proteasevaccine of the present invention, when administered to an animal in aneffective manner, is able to elicit (i.e., stimulate) an immune responsethat produces an antibody titer in the blood stream of the animalsufficient to reduce flea protease activity. Similarly, an anti-fleaprotease antibody of the present invention, when administered to ananimal in an effective manner, is administered in an amount so as to bepresent in the animal's blood stream at a titer that is sufficient toreduce flea protease activity. A protease inhibitor compound of thepresent invention, when administered to an animal in an effectivemanner, is administered in a manner so as to be present in the animal'sblood stream at a concentration that is sufficient to reduce fleaprotease activity. Oligonucleotide nucleic acid molecules of the presentinvention can also be administered in an effective manner, therebyreducing expression of flea proteases.

Compositions of the present invention can be administered to animalsprior to or during flea infestation. It is to be noted that whenvaccines of the present invention are administered to an animal, a timeperiod is required for the animal to elicit an immune response beforethe animal is competent to inhibit protease activity of fleas feedingfrom that animal. Methods to obtain an immune response in an animal areknown to those skilled in the art.

Acceptable protocols to administer compositions in an effective mannerinclude individual dose size, number of doses, frequency of doseadministration, and mode of administration. Determination of suchprotocols can be accomplished by those skilled in the art. A suitablesingle dose is a dose that is capable of protecting an animal from fleainfestation when administered one or more times over a suitable timeperiod. For example, a preferred single dose of a protease vaccine or amimetope thereof ranges from about 1 microgram (μg, also denoted ug) toabout 10 milligrams (mg) of the composition per kilogram body weight ofthe animal. Booster vaccinations can be administered from about 2 weeksto several years after the original administration. Booster vaccinationspreferably are administered when the immune response of the animalbecomes insufficient to protect the animal from flea infestation. Apreferred administration schedule is one in which from about 10 μg toabout 1 mg of the vaccine per kg body weight of the animal isadministered from about one to about two times over a time period offrom about 2 weeks to about 12 months. In one embodiment, a booster doseof a composition of the present invention is administered about 4 to 6weeks after the primary dose, and additional boosters are administeredabout once or twice a year. Modes of administration can include, but arenot limited to, oral, nasal, topical, transdermal, rectal, andparenteral routes. Parenteral routes can include, but are not limited tosubcutaneous, intradermal, intravenous, and intramuscular routes.

In another embodiment, a preferred single dose of an anti-flea proteaseantibody composition or a mimetope thereof ranges from about 1 μg toabout 10 mg of the composition per kilogram body weight of the animal.Anti-flea antibodies can be re-administered from about 1 hour to aboutbiweekly for several weeks following the original administration.Booster treatments preferably are administered when the titer ofantibodies of the animal becomes insufficient to protect the animal fromflea infestation. A preferred administration schedule is one in whichfrom about 10 μg to about 1 mg of an anti-flea protease antibodycomposition per kg body weight of the animal is administered about every2 to every 4 weeks. Suitable modes of administration are as disclosedherein and are known to those skilled in the art.

According to one embodiment, a nucleic acid molecule of the presentinvention can be administered to an animal in a fashion to enableexpression of that nucleic acid molecule into a protective protein(e.g., flea protease vaccine, anti-flea protease antibody, orproteinaceous protease inhibitor) or protective RNA (e.g., antisenseRNA, ribozyme or RNA drug) in the animal to be protected from disease.Nucleic acid molecules can be delivered to an animal in a variety ofmethods including, but not limited to, (a) direct injection (e.g., as"naked" DNA or RNA molecules, such as is taught, for example in Wolff etal., 1990, Science 247, 1465-1468) or (b) packaged as a recombinantvirus particle vaccine or as a recombinant cell vaccine (i.e., deliveredto a cell by a vehicle selected from the group consisting of arecombinant virus particle vaccine and a recombinant cell vaccine).

A recombinant virus particle vaccine of the present invention includes arecombinant molecule of the present invention that is packaged in aviral coat and that can be expressed in an animal after administration.Preferably, the recombinant molecule is packaging-deficient. A number ofrecombinant virus particles can be used, including, but not limited to,those based on alphaviruses, poxviruses, adenoviruses, herpesviruses,and retroviruses.

When administered to an animal, a recombinant virus particle vaccine ofthe present invention infects cells within the immunized animal anddirects the production of a protective protein or RNA nucleic acidmolecule that is capable of protecting the animal from disease caused bya parasite of the present invention. A preferred single dose of arecombinant virus particle vaccine of the present invention is fromabout 1×10⁴ to about 1×10⁷ virus plaque forming units (pfu) per kilogrambody weight of the animal. Administration protocols are similar to thosedescribed herein for protein-based vaccines.

A recombinant cell vaccine of the present invention includes recombinantcells of the present invention that express at least one protein of thepresent invention. Preferred recombinant cells include Salmonella, E.coli, Mycobacterium, S. frugiperda, baby hamster kidney, myoblast G8,COS, MDCK and CRFK recombinant cells, with Salmonella recombinant cellsbeing more preferred. Such recombinant cells can be administered in avariety of ways but have the advantage that they can be administeredorally, preferably at doses ranging from about 10⁸ to about 10¹²bacteria per kilogram body weight. Administration protocols are similarto those described herein for protein-based vaccines. Recombinant cellvaccines can comprise whole cells or cell lysates.

Compositions of the present invention can be administered to any animalsusceptible to flea infestation, including warm-blooded animals.Preferred animals to treat include mammals and birds, with cats, dogs,humans, cattle, chinchillas, ferrets, goats, mice, minks, rabbits,raccoons, rats, sheep, squirrels, swine, chickens, ostriches, quail andturkeys as well as other furry animals, pets and/or economic foodanimals, being more preferred. Particularly preferred animals to protectare cats and dogs.

The present invention includes compositions to treat flea infestation byany flea. As such, compositions of the present invention can be derivedfrom any flea species. Preferred fleas to target include fleas of thefollowing genera: Ctenocephalides, Cyopsyllus, Diamanus (Oropsylla),Echidnophaga, Nosopsyllus, Pulex, Tunga, and Xenopsylla, with those ofthe species Ctenocephalides canis, Ctenocephalides felis, Diamanusmontanus, Echidnophaga gallinacea, Nosopsyllus faciatus, Pulex irritans,Pulex simulans, Tunga penetrans and Xenopsylla cheopis being morepreferred. Particularly preferred fleas from which to protect animalsinclude fleas of the species Ctenocephalides felis, Ctenocephalidescanis, and Pulex species (e.g., Pulex irritans and Pulex simulans). Itis also within the scope of the present invention to administercompositions of the present invention directly to fleas.

The present invention also includes the use of compositions of thepresent invention to reduce infestation by other ectoparasites as wellas the use of compositions including protease vaccines, anti-proteaseantibodies and compounds that inhibit protease synthesis and/or activityderived from any ectoparasite to reduce ectoparasite infestation,particularly controlled release formulations containing suchcompositions. Preferred ectoparasites to target include arachnids,insects and leeches. More preferred ectoparasites to target includefleas; ticks, including both hard ticks of the family Ixodidae (e.g.,Ixodes and Amblyomma) and soft ticks of the family Argasidae (e.g.,Ornithodoros, such as O. parkeri and O. turicata); flies, such as midges(e.g., Culicoides), mosquitos, sand flies, black flies, horse flies,horn flies, deer flies, tsetse flies, stable flies, myiasis-causingflies and biting gnats; ants; spiders, lice; mites; and true bugs, suchas bed bugs and kissing bugs, including those carrying Chagas disease.Even more preferred ectoparasites to target include fleas, mosquitos,midges, sandflies, blackflies, ticks and Rhodnius.

The following examples are provided for the purposes of illustration andare not intended to limit the scope of the present invention.

EXAMPLES Example 1

This Example demonstrates that soluble flea midgut preparations containserine protease activity as well as leucine aminopeptidase activity.

Using a homogenization/sonication protocol as described in U.S. Pat. No.5,356,622, ibid., followed by an about 2 minute centrifugation step atabout 10,000×g, soluble flea midgut preparations were obtained from fedand unfed fleas. Pellets from the centrifugation step were alsocollected and resuspended for analysis. Also prepared were whole flealysates. Peptide substrate screening studies using the syntheticchromogenic trypsin substrate BAPNA(Nα-benzoyl-DL-arginine-p-nitroanilide; available from Sigma ChemicalCo., St. Louis Mo.) demonstrated significant proteolytic activity inboth soluble flea midgut preparations as well as some proteolyticactivity in resuspended midgut pellets. Soluble unfed flea midgutpreparations exhibited about 10 times as much activity as did controls(samples to which no flea midgut fractions were added), whereas solublefed flea midgut preparations exhibited about 20 times as much activityas did controls. Whole flea preparations exhibited about 2 to 3 times asmuch activity as did controls.

The ability of soluble fed and unfed flea midgut preparations to cleaveBAPNA was almost completely inhibited (i.e., nearly 100%) by aprotinin(available from Sigma), whereas PMSF (phenylmethane-7-sulfonyl fluoride;available from Sigma) inhibited such proteolytic activity by about 50%.EDTA inhibited proteolytic activity of the preparations by about 10%,whereas addition of calcium ions stimulated proteolytic activity byabout 25%. These results indicate the presence of serine proteaseactivity, and more particularly of trypsin-like activity, in thesesoluble flea midgut preparations. These results also suggest thepresence of serine protease isoforms in the preparations. It is also ofinterest to note that flea trypsin-like activity appears to bedistinctive from that of mosquitos in that mosquito trypsins are notaffected by EDTA or calcium ions.

Using a methyl-hemoglobin substrate, the pH optimum of the proteolyticactivity in the soluble flea midgut preparations was found to be betweenpH 7 and pH 9, with a pH of about pH 8 giving the best activity. Such pHoptima suggest the presence of serine proteases in soluble flea midgutpreparations.

Soluble preparations of both unfed and fed flea midgut solublepreparations also were able to cleave the leucine aminopeptidasespecific substrate LPNA (L-leucine-p-nitroanilide; available from Sigma)using standard conditions, indicating the presence of leucineaminopeptidase (LAP) activity in such preparations.

Example 2

The following example evaluated the number of proteases in flea midgutsthat could be assessed by protease substrate gel analysis.

Protease substrate gels (available from Novex, San Diego, Calif., asNovex Zymogels) were 10% polyacrylamide-SDS gels with 0.1% gelatin.Samples and gels were processed according to Novex instructions.Briefly, samples were diluted in SDS-PAGE sample buffer without reducingagents. Tris-glycine SDS-PAGE was carried out by standard procedures.After electrophoresis, gels were incubated in 0.25% Triton X-100 at roomtemperature for 30 minutes (min), then in developing buffer (50 mM(millimolar) Tris-HCl pH 7.0, 5 mM CaCl₂, 0.02% Brij 35, 0.2 M (molar)NaCl) at room temperature for 30 min, and then incubated with freshdeveloping buffer at 37° C., usually overnight. Gels were then stained30 min in 0.5% coomassie R-250, 40% methanol, 10% acetic acid anddestained in 40% methanol, 10% acetic acid.

The following flea midguts were dissected directly into sample buffer:100 midguts from unfed males; 100 midguts from unfed females; 100midguts from fed males; and 100 midguts from fed females. Samplescontaining 10 or 20 midguts each were evaluated using protease substrategel analysis and numerous negative staining bands were observed. Thegeneral pattern was the same for female and male midguts, although thereappeared to be more activity in gel lanes containing female midguts.There were distinct differences noted between gel lanes containing fedand unfed midguts. There was a definite increase in overall activity inthe fed midgut lanes, and, in addition, there were differences in theband patterns.

Fed and unfed female midguts were further evaluated using proteasesubstrate gel analysis and the results are shown in FIG. 1. The proteasesubstrate gel shown in FIG. 1 demonstrates the relative proteolyticactivity in 1, 2, 5 or 10 midguts from either fed or unfed female fleas.Specifically, lane 1 contains a set of molecular weight markers. Lanes 2through 5 contain, respectively, 10, 5, 2 and 1 unfed midguts. Lanes 6through 9 contain, respectively, 1, 2, 5 and 10 fed midguts. Lane 10contains 100 μg of dried bovine blood.

Proteolytic activity could easily be detected in one fed or one unfedfemale midgut, although there was considerably more activity in the fedmidgut. Lane 10 evaluated 100 μg of dried bovine blood to assess if theincrease in activity seen in the fed midgut lane was due to proteases inthe blood meal. No activity was seen in the blood lane.

Example 3

This example evaluated the protease classes present in flea midguts.

Three unfed female midguts and 0.75 fed female midguts were evaluated induplicate in several protease substrate gels. Each gel was cut in half.Half was processed as described in Example 2, while the other halfcontained protease inhibitors in all incubation buffers. The followinginhibitors were evaluated:

(a) the serine protease inhibitor AEBSF (available from BoehringerMannheim, Indianapolis, Ind.) was used at a final concentration of 1 mM;

(b) the serine protease inhibitor soybean trypsin inhibitor (availablefrom Sigma) was used at a final concentration of 100 μg/ml (milliliter);

(c) the cysteine and serine protease inhibitor leupeptin (available fromSigma) was used at a final concentration of 10 μg/ml;

(d) the aminopeptidase inhibitor bestatin (available from Sigma) wasused at a final concentration of 0.25 mM;

(e) the metalloprotease inhibitor EDTA (available from Sigma) was usedat a final concentration of 2 mM; and

(f) the cysteine protease E-64 (available from Sigma) was used at afinal concentration of 10 μg/ml.

AEBSF, soybean trypsin inhibitor and leupeptin were the only inhibitorsto have any effect at the sensitivity of this assay. It was determinedthat serine proteases were the predominant, if not only, proteasespresent in the midgut preparations evaluated. FIG. 2 shows a proteasesubstrate gel with fed (lanes 2 and 4) and unfed (lanes 1 and 3) midgutpreparations with (lanes 3 and 4) and without (lanes 1 and 2) AEBSF.Residual activity in the inhibitor lanes could have been due toproteolysis that occurred during electrophoresis and prior to saturationof the gel with inhibitor in the incubation buffers.

Example 4

This Example evaluates protease activity contained in a soluble fedmidgut preparation of the present invention.

Mixed-sex fed flea midguts were processed as described in U.S. Pat. No.5,356,622, ibid. Aliquots of several steps of the procedure wereevaluated by loading an equivalent of 0.4 midguts per lane of a proteasesubstrate gel as described in Example 2. The results are shown in FIG.3. Samples were from the low speed supernatant (lanes 2 and 9),sonicated midguts (lanes 3 and 10), high speed supernatant (lanes 4 and11), combined low and high speed supernatants (FGS) (lanes 5 and 12) andthe high speed pellet (lanes 6 and 13). Lanes 7 and 8 contained 50nanograms (ng) of trypsin as a control. Duplicate lanes were evaluated.The gel was cut in half, and lanes 1-7 were processed as described inExample 2, and lanes 8-14 were processed with 100 μg/ml soybean trypsininhibitor in all the incubation buffers.

Protease activity was seen in all preparations, the most being observedin the FGS lane (lane 5). It was also evident that the majority of theactivity was inhibited by soybean trypsin inhibitor, a serine proteaseinhibitor.

Example 5

This Example demonstrates the increase in flea midgut protease activityafter blood feeding by fleas.

Fleas were fed on a dog for 15 minutes. At timed intervals afterfeeding, two midguts were dissected directly into sample buffer andproteases evaluated by protease substrate gel analysis as described inExample 2. FIG. 4 depicts a gel showing midgut protease activity at 30min (lane 1), 1 hr (lane 2), 2 hr (lane 3), 4 hr (lane 4), 6 hr (lane5), 8 hr (lane 6), 24 hr (lane 7) and 56 hr (lane 8) after blood feedingended.

Increases in proteolytic activity were first observed 2 hr (lane 3)after feeding, although at 4 hr (lane 4) there was a much greaterincrease in activity noted. This increase in activity was still noticed56 hr after feeding (lane 8).

Example 6

This Example evaluates the effect of a number of protease inhibitors onflea viability and fecundity in a flea feeding system as described byWade et al. ibid.

The following protease inhibitors were tested at the indicated finalconcentrations in blood meals:

(a) Aminopeptidase inhibitor bestatin at 1.3 mM and 13 mM;

(b) Aspartic acid protease inhibitor pepstatin A at 1 μg/ml and 10μg/ml;

c) Cysteine protease inhibitor E-64 at 1 μg/ml and 10 μg/ml.

(d) Metalloprotease inhibitor phosphoramidon at 10 μAg/ml and 100 μg/ml;and

(e) the following serine protease inhibitors:

AEBSF at 0.3 mM, 0.5 mM, 5.0 mM and 6.0 mM;

Aprotinin at 2 μg/ml and 20 μg/ml;

Leupeptin at 5 μg/ml and 50 μg/ml;

Soybean trypsin inhibitor at 10 μg/ml and 100 μg/ml;

Soybean trypsin/chymotrypsin inhibitor at 10 μg/ml and 100 μg/ml;

AEBSF is available from Boehringer Mannheim; all other listed inhibitorsare available from Sigma.

Protease inhibitor compounds were tested in groups of 3 to 6 includingappropriate control groups. Inhibitors were not tested in groups ofcommon inhibition types. Rather, they were tested in groups based on thediluent needed to dissolve them. (AEBSF, aprotinin, bestatin, leupeptin,phosphoramidon, soybean trypsin inhibitor and soybeantrypsin/chymotrypsin inhibitor were dissolved in water; E-64 andpepstatin were dissolved in ethanol). This reduced the number of control(diluent only) groups needed within a particular assay. Inhibitorconcentrations were chosen such that the lower concentration used waswithin the range recommended by the supplier for that inhibitor. Thehigher concentration was typically 10 times above the lowerconcentration and was used to look for dose response.

The general protocol for all of the assays was as follows: Approximately2000 newly emerged adult fleas were placed in feeding chambers to feedon normal blood for about 24 to 48 hr. The fleas were prefed for tworeasons: The first was to be certain that only fleas that would feed inthe feeding system were used in the comparative study. The second was toprime female fleas for egg laying, since female fleas typically do notbegin laying maximal numbers of eggs per day until the third day offeeding.

The prefed fleas were placed in "minifeeder" feeding chambers at a ratioof about 80 female fleas to about 20 male fleas for a total of about 100fleas per chamber. Actual total number of fleas per chamber varied fromabout 90 to 125 fleas. Previous experiments have not demonstrated anydifferences in adult survival or fecundity based on such variance innumbers of fleas in a chamber. Three chambers were prepared for eachexperimental and control group. A fresh blood meal containing theappropriate inhibitor in 3 ml total volume was placed on each chamberdaily through the 7 day extent of an assay.

On days 3, 5, and 7 of the assay, surviving adult fleas were transferredto clean chambers. The contents of the original chambers were dissolvedin about 40 ml of PBS (phosphate-buffered saline) in a 50 ml Falcontube. The contents of a given tube were then filtered through apre-weighed #1 Whatman filter disk inserted into a vacuum filter. The 50ml tube and the filter funnel were rinsed with distilled water which wasthen passed through the filter. Once the chamber contents had beenfiltered, dead adult fleas were removed from the filter paper and placedin a labelled tube so that they could be counted and sexed. The filterpaper was then placed into a preweighed 12×75 polypropylene tube anddried in the SpeedVac for 2.5 hr with the heater on. After drying thefilter paper was weighed. The weight of the filter paper and tube wassubtracted to obtain the dry weight of the eggs and this value wasconverted to an estimated number of eggs using the formulay=41384.361×+162.37, where x=dry weight of eggs.

On day 7, adult fleas that had survived the study were frozen, countedand sexed. The numbers were added to the number of male and female fleasthat had died during the assay to verify the number of male and femalefleas in each chamber at the start of the study.

Female, male and total adult flea survival were calculated for allexperimental and control groups on days 3, 5, and 7 of each assay.Additionally, the number of eggs per surviving female was calculated ondays 3, 5 and 7. Female fleas found dead on a given collection date wereincluded in the total number of egg-laying females for the days betweenthat date and the previous collection date, providing a conservativeestimate of fecundity. Fecundity values were averaged for the threecollection dates to obtain an average for each group over 7 days.

Results of these studies are presented below in Table 1 and FIG. 6through FIG. 9. All survival and fecundity values are presented below asa percent of control value.

                  TABLE 1                                                         ______________________________________                                        Effect of Protease Inhibitors                                                   on Flea Viability and Fecundity                                                                    Fecundity.sup.1                                                                       Adult Survival.sup.1                           Compound  Conc.     Days 1-7 Female                                                                              Male  Total                                ______________________________________                                        AEBSF     6.0    mM     17.2%  4.1%  0.0   3.4%                                  5.0 mM 1.4% 6.8% 0.0% 5.6%                                                    0.5 mM 95.0% 103.9% 104.2% 103.6%                                             0.3 mM 82.4% 116.2% 103.0% 111.9%                                            Aprotinin 20 ug/ml 84.2% 100.0% 101.7% 99.9%                                   2 ug/ml 83.2% 103.2% 104.9% 103.3%                                           Leupeptin 50 ug/ml 77.6% 101.5% 111.7% 104.6%                                  5 ug/ml 85.0% 71.0% 61.4% 68.4%                                              Soybean Trypsin 100 ug/ml 79.1% 76.5% 76.0% 76.3%                             Inhibitor 10 ug/ml 96.1% 80.1% 101.7% 83.9%                                   Trypsin/ 100 ug/ml 81.1% 88.0% 95.4% 89.9%                                    Chymotrypsin 10 ug/ml 100.7% 115.1% 143.5% 120.7%                             Inhibitor                                                                     E-64 10 ug/ml 177.4% 110.2% 139.0% 114.2%                                      1 ug/ml 109.4% 99.9% 102.9% 100.1%                                            10 ug/ml 84.1% 90.2% 91.1% 90.6%                                              1 ug/ml 95.2% 77.3% 80.0% 77.5%                                              Phosphoramidon 100 ug/ml 84.9% 70.2% 64.6% 69.7%                               10 ug/ml 89.0% 98.8% 95.2% 97.8%                                             Pepstatin A 10 ug/ml 83.9% 113.6% 133.4% 116.2%                                1 ug/ml 67.7% 77.6% 96.6% 80.5%                                              Bestatin 13.0 mM 23.3% 121.0% 103.4% 117.0%                                    1.3 mM 60.4% 119.5% 116.3% 116.8%                                          ______________________________________                                         .sup.1 All experimental values are expressed as a percent of the              corresponding control group.                                             

The aminopeptidase inhibitor bestatin caused a significant (p<0.05)reduction in fecundity at 13 mM (77% reduction) and at 1.3 mM (40%reduction) indicating the presence of an aminopeptidase or otherexopeptidase in flea midguts. Bestatin at the concentrations tested,however, had no significant effect on adult viability at eitherconcentration. These results suggest that aminopeptidases may play arole in ovarian function, or a related process, such as vitellogenesis.

The aspartic acid protease inhibitor pepstatin A caused a significantreduction (p<0.05) in fecundity at 1 μg/ml (32% reduction), but not at10 μg/ml. Pepstatin A had no significant effect on adult viability ateither concentration.

The cysteine protease inhibitor E-64 showed no statistically significantreduction in fecundity in this assay. There was a small, but significant(p<0.05), reduction in total adult flea survival when E-64 was dissolvedin grain alcohol and added to blood at 1 μg/ml. However, this reductionwas not evident in the group that was fed blood containing 10 μg/ml E-64in grain alcohol.

The metalloprotease inhibitor phosphoramidon caused a reduction in adultviability of about 30%, which, however was not statisticallysignificant. There was no significant reduction in fecundity.

Results using serine protease inhibitors were particularly interestingand suggest the significance of serine proteases in flea midguts. AEBSFadministered at concentrations ranging from about 5 mM to about 6 mMreduced flea fecundity by more than 80%. In addition, adult survival wasreduced to near zero (p<0.05).

Aprotinin, however, had no significant effect on either fecundity orviability, likely due to the ability of serum proteins, such as albumin,to interfere with aprotinin's inhibitory activity.

Leupeptin had no effect on fecundity at both concentrations, but reducedadult viability by 30% at 5 μg/ml. However, adult viability was notaffected by 50 μg/ml leupeptin and none of the observed reductions werestatistically significant.

Soybean trypsin inhibitor caused a small (20%) statisticallyinsignificant reduction in fecundity at 100 μg/ml. The lowerconcentration had no effect. Soybean trypsin inhibitor, on the otherhand, is very effective in in vitro studies as disclosed in several ofthe examples and was used to purify serine proteases as disclosed inExample 7. Soybean trypsin/chymotrypsin inhibitor had no effect on adultviability or fecundity.

Example 7

This Example describes the production of a preferred soluble flea midgutpreparation of the present invention and purification of flea serineproteases therefrom. Also included is amino acid sequence analysis of aflea serine protease of the present invention.

The soluble flea midgut preparation was prepared as follows. Fleamidguts (3,735) from a mix of female and male fed fleas were homogenizedin a homogenization buffer comprising 1.5 ml 50 mM Tris-HCl, 0.5 M NaCl,pH 8.5. The homogenate was centrifuged at 14,000×g for 10 min. Theresultant pellet was processed again in another 1.5 ml of thehomogenization. The two supernatant solutions were combined to form thesoluble flea midgut preparation.

The preparation was added to 3 ml of p-aminobenzamidine-sepharose 6 B(affinity matrix for trypsin-like proteases, available from Sigma) andincubated at 5° C. overnight on a rocker. The sepharose beads weredrained and washed with 7.5 ml of the homogenization buffer. Theadsorbed proteins were eluted with 5 ml 0.1 M p-aminobenzamidine in thesame buffer. This eluate was concentrated and the buffer exchanged to 50mM Tris-HCl pH 8.5, 0.1 mM CaCl₂ by ultrafiltration through a membranewith a 3 kD cutoff, the final volume being 140 μl (microliters).

Labeling of proteins was performed by adding 10 μl of (1, 3-³H)-diisopropylfluorophosphate (available from New England Nuclear,Beverly, Mass. at 6.0 Ci (Curies)/mmole, 1.0 mCi/ml) to 90 μl of theaffinity purified proteins and incubating at 5° C. for 18 hours. Thereaction was divided in half, each half then being separated by C4reverse phase chromatography according to the following protocol:

Buffer A: 0.1% TFA in water

Buffer B: 0.085% TFA, 90% Acetonitrile

0.8 ml/min, 220 nm, 1 min fractions

5.6% B 15 min

5.6% to 100% B over 60 min

Ten microliters of each fraction was added to scintillation fluid andcounted. Most protein-associated counts were found in fractions 44-47.FIG. 5A shows electrophoresis of fractions 40 (lane 2), 44 (lane 3), 46(lane 4) and 47 (lane 5) from one chromatography run through a 14%Tris-glycine polyacrylamide-SDS gel, followed by coomassie staining.This gel was then processed with Entensigy (NEN) and exposed to film for18 hours, as shown in FIG. 5B. Each fraction contained several proteinsas shown in FIG. 5A, but only 4 bands were labeled, the most prominentbeing 26 kd (seen in lanes 3, 4 and 5), and denoted herein as PfSP26(now denoted PafSP-26K). A faint band of 24 kd, denoted herein as PfSP24(now denoted PafSP-24K), was also noticed in lane 5. A band of 19 kD,denoted herein as PfSP19 (now denoted PafSP-19K), was labeled in lane 4that was associated with a very faintly staining protein band. Somelabeled proteins were seen at the dye front of lanes 4 and 5, indicatinga molecular weight less than 6 kd, denoted herein as PfSP6 (now denotedPafSP-6K), and could be degradation products.

Fraction 44 (analogous to lane 3) from a second C4 chromatographyseparation experiment was electrophoresed, blotted onto PVDF, stainedwith Coomassie R-250 and destained via standard procedures. The 26 kdband, corresponding to PfSP26 (also referred to herein as PfSP44-E,indicating the fraction in which the protein eluted and the gel/filterband from which the protein was excised), was excised and subjected toN-terminal amino acid sequencing using techniques known to those skilledin the art. A partial N-terminal amino acid sequence of about 32 aminoacids was deduced and is represented herein as SEQ ID NO:1:

I I G G E V A G E G S A P Y Q V S L R T K E G N H F S G G S I L,

It should be noted that since amino acid sequencing technology is notentirely error-free, SEQ ID NO:1 represents, at best, an apparentpartial N-terminal amino acid sequence of PfSP26. This caution isparticularly relevant in light of the sequencing of this protein havingbeen done at a low picomolar concentration.

A homology search of the non-redundant protein sequence database wasperformed through the National Center for Biotechnology Informationusing the BLAST network. This database includes +SwissProt +PIR+SPUpdate +GenPept + GPUpdate.ole level. Results of the search indicatethat the N-terminus of PfSP26 shares significant amino sequence homologywith a number of serine proteases, including a variety of trypsins,chymotrypsins and plasmins. The 32-amino acid N-terminal amino acidsequence of PfSP26 shared the highest degree of homology with a hornetchymotrypsin II.

Example 8

This example describes the cloning of certain flea protease nucleic acidmolecules of the present invention. This example also describes theproduction of certain recombinant molecules, recombinant cells and fleaprotease proteins of the present invention.

Several flea serine protease nucleic acid molecules, ranging in sizefrom about 250 to about 500 nucleotides, and representing one or morepartial flea serine protease genes, were PCR amplified from a fed fleamidgut cDNA library that was prepared from RNA isolated from fed fleamidguts using standard protocols as described in Sambrook et al., ibid.Several pairs of primers were used in PCR amplification reactions thatrepresented degenerate oligonucleotides designed from publishedsequences of serine protease genes isolated from biting insects (e.g.,mosquitos and black flies). Each primer pair was designed so that aproperly amplified fragment of a flea serine protease gene would includea domain corresponding to the most conserved domain of trypsin proteasegenes (thought to be the active site) given that such a domain iscontained in flea serine protease gene(s).

The amplified PCR fragments were of predicted size, ranging from about250 nucleotides to about 500 nucleotides, depending on which primerpairs were used. PCR fragments that hybridized to a probe designed fromthe domain most conserved among all known trypsin genes were gelpurified and cloned, for example, into the PCRII cloning vector(available from InVitrogen, Corp., San Diego, Calif.), followingmanufacturer's instructions. Nucleic acid sequences of the fragments arebeing determined using standard techniques.

The amplified PCR fragments are also being used as probes to identifyfull-length flea protease genes in unfed and fed flea midgut cDNAlibraries and in flea salivary gland cDNA libraries, as well as in fleagenomic DNA libraries, using standard procedures.

Recombinant molecules and recombinant cells including the amplified PCRfragments as well as full-length flea protease genes are being producedusing standard procedures. Culturing of such recombinant cells leads tothe production of flea protease proteins of the present invention.

Example 9

This Example describes the testing of a flea protease protein as a fleaprotease vaccine of the present invention, that is for the ability ofsuch a protein, upon administration to an animal, to elicit theproduction of antibodies that reduce flea protease activity and, assuch, reduce flea viability and/or fecundity. This Example alsodemonstrates the use of such a flea protease protein as a vaccine on adog subsequently infested with fleas.

A flea protease protein produced as described in Example 7 isadministered to rabbits according to a standard immunization protocolknown to those skilled in the art, including appropriate booster shots.Such a protein is also administered to guinea pigs and to dogs followinga similar protocol.

Sera is collected from the treated rabbits and is verified to containanti-flea protease antibodies. Such sera is then fed to fleas in afeeding system as reported by Wade et al. ibid. Fleas feeding on such asera show reduced viability compared to fleas feeding on sera collectedfrom rabbits not administered the flea protease protein. Sera fromtreated guinea pigs and dogs are verified in a similar manner.

Dogs treated with a flea protease protein are then infested with fleasas are dogs not treated with a flea protease protein. Dogs treated witha flea protease protein show a significant reduction in flea burdencompared to untreated dogs.

Example 10

This Example describes the determination of the partial N-terminal aminoacid sequence of additional flea serine protease proteins of the presentinvention.

An additional eight flea serine proteases were purified and consensuspartial N-terminal amino acid sequences were determined as described inExample 7. The results are as follows, the proteins being named by thefraction in which they were eluted and the SDS-PAGE gel band from whichthey were excised. Each of the proteases bore at least some sequencehomology to known proteases, the highest percent identity estimated tobe no more than about 30-40%.

Flea protease PfSP45-C had a partial N-terminal amino acid sequence of XV G G H D T S I D X H P H Q V T, also represented herein as SEQ ID NO:2.PfSP45-C was most similar in amino acid sequence to a fruit fly trypsinepsilon.

Flea protease PfSP46-C had a partial N-terminal amino acid sequence of IV G G A D A A P G N A P F Q V S L R D K G, also represented herein asSEQ ID NO:3. PfSP46-C was most similar in amino acid sequence to acollagenolytic 36 kD protease from a Kamchatda crab.

Flea protease PfSP46-A had a partial N-terminal amino acid sequence of IV G G Q D A D I A K Y G Y Q A S L Q V F N E H F X G A X I L N N Y, alsorepresented herein as SEQ ID NO:4. PfSP46-A was most similar in aminoacid sequence to a hornet chymotrypsin II.

Flea protease PfSP46-B had a partial N-terminal amino acid sequence of IV G G T D V N I E N F G W Q V S L F D R N G H F, also represented hereinas SEQ ID NO:5. PfSP46-B was most similar in amino acid sequence to afruit fly trypsin beta.

Flea protease PfSP48-A had a partial N-terminal amino acid sequence of IV G G H D T S I D K H P F Q V S L I D K N, also represented herein asSEQ ID NO:6. PfSP48-A was most similar in amino acid sequence to a fruitfly trypsin epsilon.

Flea protease PfSP48-B had a partial N-terminal amino acid sequence of VV G G L E A A E G S A P Y Q V X L Q W G N F, also represented herein asSEQ ID NO:7. PfSP48-B was most similar in amino acid sequence to a humanFactor 12.

Flea protease PfSP48-D had a partial N-terminal amino acid sequence of IV G G E D A E L G E X P T Q, also represented herein as SEQ ID NO:8.PfSP48-D was most similar in amino acid sequence to a bovine Factor 9.

Flea protease PfSP40-B had a partial N-terminal amino acid sequence of DE D G K D D S A P G E I, also represented herein as SEQ ID NO:9.PfSP40-B was most similar in amino acid sequence to a fruit flyfurin-like protease I.

Example 11

This Example describes the isolation of nucleic acid molecules encodingflea serine protease proteins of the present invention.

Several midgut proteinase cDNA genes have been isolated in a mannersimilar to that described in Example 8, using two degenerate primers,the design of which was based on a highly conserved serine proteinaseamino acid sequence (C Q/N G D S G G P L, denoted SEQ ID NO:10) locatedabout 195 amino acid residues (based on an average protease size ofabout 240 residues) from the mature amino terminus in a number of knownserine proteases. Complementing primers for use in the PCR amplificationreaction were primers corresponding to the vectors in which nucleic acidmolecules of the present invention had been ligated. The actual primersused in PCR amplification of serine protease nucleic acid molecules fromwhole fed flea cDNA expression libraries (produced as described inExample 8) included the following serine protease specific primers:cat-try #1 having nucleic acid sequence 5' TAA WGG WCC WCC YGA ATC TCCCTG GCA 3' (Y indicating C or T; W indicating A or T), representedherein as SEQ ID NO:11; and cat-try #2 having nucleic acid sequence 5'TAA WGG WCC AGA RTC TCC TTG ACA 3' (R indicating A or G), representedherein as SEQ ID NO:12. Vector specific primers included: M13 Reversehaving nucleic acid sequence 5' GGAAACAGCTATGACCATG 3', representedherein as SEQ ID NO:13; and T3 Primer having nucleic acid sequence 5'ATTAACCCTCACTAAAG 3', represented herein as SEQ ID NO:14. The resultantPCR products, obtained using standard PCR conditions (e.g., Sambrook etal., ibid.), were about 600 to about 700 nucleotides in length.

The PCR products were hybridized under standard hybridization conditions(e.g., Sambrook et al., ibid.) with (i.e., to) an internal syntheticoligonucleotide probe named H57, the sequence of which corresponds to aregion including a conserved histidine residue in known serineproteases. The nucleic acid sequence of H57 is 5' TGG GTW GTW ACW GCWGCW CAT TG 3', represented herein as SEQ ID NO:15. PCR products whichhybridized strongly to the probe were gel purified and cloned into theTA Vector™ (available from InVitrogen, Corp.). Approximately 80recombinant TA vector clones were isolated.

To prevent repetitive sequencing of the same serine proteinase clones, anumber of the clones were characterized to identify those having uniquerestriction endonuclease patterns using the enzymes HaeII and HaeIII.About 11 plasmids apparently containing unique flea serine proteinasenucleic acid molecules of about 600 to about 700 nucleotides in lengthwere isolated using this procedure. These nucleic acid molecules weresubjected to nucleic acid sequencing using the Sanger dideoxy chaintermination method, as described in Sambrook et al., ibid.

The complete nucleic acid sequence of one of the flea serine proteasenucleic molecules, namely nfSP4₆₇₂ is represented herein as SEQ IDNO:16. Translation of SEQ ID NO:16 yields a protein of about 223 aminoacids, denoted PfSP4₂₂₃, having amino acid sequence SEQ ID NO:17.Although the entire amino acid sequence of PfSP4₂₂₃ is not highlyconserved to that of known serine proteases, there are several conservedregions of note (as numbered for SEQ ID NO:17), including: (a) thesequence IVGG spanning from about amino acid 5 through about amino acid9; (b) the active-site histidine at about amino acid 46 and surroundingsequences spanning from about amino acid 41 through about amino acid 47;(c) the conserved aspartic acid residue at about amino acid 90; (d) theGWG sequence spanning from about amino acid 124 through about amino acid126; the conserved cysteines at about amino acid 152 and about aminoacid 165; and the conserved sequence around the active site serine,spanning from about amino acid 174 through about amino acid 182.

Nucleic acid and amino acid sequences of all 11 flea serine proteasenucleic acid molecules were determined for the regions corresponding tothe region in known serine proteases to span from the conserved GWGsequence to the conserved CXGDSGGP sequence (denoted SEQ ID NO:10). Fleanucleic acid molecule nfSP1₁₅₆ has the nucleic acid sequence representedherein as SEQ ID NO:18, which encodes a protein PfSP1₅₂ having an aminoacid sequence represented herein as SEQ ID NO:19. Flea nucleic acidmolecule nfSP2₁₆₈ has the nucleic acid sequence represented herein asSEQ ID NO:20, which encodes a protein PfSP2₅₆ having an amino acidsequence represented herein as SEQ ID NO:21. Flea nucleic acid moleculenfSP3₁₇₇ has the nucleic acid sequence represented herein as SEQ IDNO:22, which encodes a protein PfSP3₅₉ having an amino acid sequencerepresented herein as SEQ ID NO:23. Flea nucleic acid molecule nfSP4₁₅₆has the nucleic acid sequence represented herein as SEQ ID NO:24, whichencodes a protein PfSP4₅₂ having an amino acid sequence representedherein as SEQ ID NO:25. Flea nucleic acid molecule nfSP5₁₅₉ has thenucleic acid sequence represented herein as SEQ ID NO:26, which encodesa protein PfSP5₅₃ having an amino acid sequence represented herein asSEQ ID NO:27. Flea nucleic acid molecule nfSP6₁₆₈ has the nucleic acidsequence represented herein as SEQ ID NO:28, which encodes a proteinPfSP6₅₆ having an amino acid sequence represented herein as SEQ IDNO:29. Flea nucleic acid molecule nfSP7₁₅₉ has the nucleic acid sequencerepresented herein as SEQ ID NO:30, which encodes a protein PfSP7₅₃having an amino acid sequence represented herein as SEQ ID NO:31. Fleanucleic acid molecule nfSP8₁₈₆ has the nucleic acid sequence representedherein as SEQ ID NO:32, which encodes a protein PfSP8₆₂ having an aminoacid sequence represented herein as SEQ ID NO:33. Flea nucleic acidmolecule nfSP⁹ ₁₆₈ has the nucleic acid sequence represented herein asSEQ ID NO:34, which encodes a protein PfSP9₅₆ having an amino acidsequence represented herein as SEQ ID NO:35. Flea nucleic acid moleculenfSP10₁₂₀ has the nucleic acid sequence represented herein as SEQ IDNO:36, which encodes a protein PfSP10₄₀ having an amino acid sequencerepresented herein as SEQ ID NO:37. Flea nucleic acid molecule nfSP11₁₆₂has the nucleic acid sequence represented herein as SEQ ID NO:38, whichencodes a protein PfSP11₅₄ having an amino acid sequence representedherein as SEQ ID NO:39.

Comparison of the nucleic acid sequences of the flea serine proteaseswith that of a mosquito (A. aegypti) trypsin indicates that SEQ ID NO:18is about 33% identical, SEQ ID NO:20 is about 33% identical, SEQ IDNO:22 is about 24% identical, SEQ ID NO:24 is about 25% identical, SEQID NO:26 is about 32% identical, SEQ ID NO:28 is about 38% identical,SEQ ID NO:30 is about 33% identical, SEQ ID NO:32 is about 33%identical, SEQ ID NO:34 is about 40% identical, SEQ ID NO:36 is about33% identical, and SEQ ID NO:38 is about 29% identical, to thecorresponding region of the mosquito trypsin. Comparison of the nucleicacid sequences of the flea serine proteases with that of a black fly (S.vittatum) trypsin indicates that SEQ ID NO:18 is about 34% identical,SEQ ID NO:20 is about 34% identical, SEQ ID NO:22 is about 25%identical, SEQ ID NO:24 is about 28% identical, SEQ ID NO:26 is about36% identical, SEQ ID NO:28 is about 45% identical, SEQ ID NO:30 isabout 29% identical, SEQ ID NO:32 is about 36% identical, SEQ ID NO:34is about 42% identical, SEQ ID NO:36 is about 34% identical, and SEQ IDNO:38 is about 30% identical, to the corresponding region of the blackfly trypsin. It is to be noted that the mosquito and black fly trypsinsare about 50% identical in the same regions.

Comparison of the amino acid sequences of the flea serine proteases withthat of a mosquito (A. aegypti) trypsin indicates that SEQ ID NO:19 isabout 11% identical, SEQ ID NO:21 is about 30% identical, SEQ ID NO:23is about 19% identical, SEQ ID NO:25 is about 19% identical, SEQ IDNO:27 is about 28% identical, SEQ ID NO:29 is about 21% identical, SEQID NO:31 is about 14% identical, SEQ ID NO:33 is about 22% identical,SEQ ID NO:35 is about 30% identical, SEQ ID NO:37 is about 22%identical, and SEQ ID NO:39 is about 29% identical, to the correspondingregion of the mosquito trypsin. Comparison of the amino acid sequencesof the flea serine proteases with that of a black fly (S. vittatum)trypsin indicates that SEQ ID NO:19 is about 14% identical, SEQ ID NO:21is about 28% identical, SEQ ID NO:23 is about 16% identical, SEQ IDNO:25 is about 17% identical, SEQ ID NO:27 is about 35% identical, SEQID NO:29 is about 33% identical, SEQ ID NO:31 is about 11% identical,SEQ ID NO:33 is about 22% identical, SEQ ID NO:35 is about 33%identical, SEQ ID NO:37 is about 21% identical, and SEQ ID NO:39 isabout 25% identical, to the corresponding region of the black flytrypsin. It is to be noted that the mosquito and black fly trypsins areabout 50% identical in the same regions.

Partial N-terminal amino acid sequences were deduced for each of thecloned flea serine protease nucleic acid molecules, four of which wereidentical to the following amino acid sequences derived from N-terminalsequencing of serine proteases as described in Example 10: SEQ ID NO:1,SEQ ID NO:4, SEQ ID NO:6 and SEQ ID NO:7. The remaining nucleic acidmolecules had the following deduced N-terminal amino acid sequences: SEQID NO:40, namely I V G G E N A K E K S D V P Y Q V S L R N A E N K H F CG G A I I D D Y W V L T, which was most similar in amino acid sequenceto mite fecal allergen Der pIII; SEQ ID NO:41, namely I V G G L E A K NG S A P F M V S L Q A E D Y F H, which was most similar in amino acidsequence to a chymotrypsin-like protein; SEQ ID NO:42, namely I I G G EV A G E G S A P Y Q V S L R T K E G N H F, which was most similar inamino acid sequence to a chymotrypsin-like protein; SEQ ID NO:43, namelyI V G G T A V D I R G F P G R Y Q F K P K P S F L W W F Y, which did notsubstantially match any protein in the data base; SEQ ID NO:44, namely IV N G L E A G V G Q F P I Q V F L D L T N I R D E K S R C G G A L F,which was most similar in amino acid sequence to a trypsin precursor;SEQ ID NO:45, namely I V G G L E A K N G I T P F I G F F A S G R L F,which was most similar in amino acid sequence to a chymotrypsin-likeprotease; SEQ ID NO:46, namely I V G G N D V S X K I F W Q V S I Q S N XQ H F C G, which was most similar in amino acid sequence to a trypsin;and SEQ ID NO:47, namely I I G G E D A P E G S A P Y Q V S L R N Q N L EH F C G G S I, which was most similar in amino acid sequence to achymotrypsin-like protein.

Additional amino terminal and carboxyl terminal sequences of flea serineprotease nucleic acid molecules comprising sequences listed above aswell as additional nucleic acid molecules identified using thetechniques described herein are presented in Table 2.

                                      TABLE 2                                     __________________________________________________________________________    Additional Flea Serine Protease Sequences                                     __________________________________________________________________________    A. The apparent N-terminal nucleic acid sequence (SEQ ID                        NO:52), as well as deduced amino acid sequence                                (SEQ ID NO:53) of nfSPl is:                                                   TCA GCA CTC GTT GCC TTG TCT GCA GCT ATT CCT CAC TCC AAC AGA                   GTC                                                                            S   A   L   V   A   L   S   A   A   I   P   H   S   N   R                    V                                                                              - GTT GGA GGA CTG GAA GCT GCA GAG GGT TCT GCA CCT TAT CAA GTA                TCC                                                                            V   G   G   L   E   A   A   E   G   S   A   P   Y   Q   V                    S                                                                             TTG CAA GTT GGC AAC TTC CAC TTC TGT GGT GGT TCA ATT CTG AAC                   GAA                                                                            L   Q   V   G   N   F   H   F   C   G   G   S   I   L   N                    E                                                                              - TAT TGG GTT TTG ACT GCT GCT CAC TGT TTG GGT TAT GAC TTC GAC                GTG                                                                            Y   W   V   L   T   A   A   H   C   L   G   Y   D   F   D                    V                                                                              - GTA GTT GGA ACA AAC AAA CTT GAT CAA CCA GGT GAA AGA TAC CTC                GTA                                                                            V   V   G   T   N   K   L   D   Q   P   G   E   R   Y   L                    V                                                                              - GAA CAA ACT TTT GTT CAC                                                     E   Q   T   F   V   H                                                         - B. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:54), as well as deduced amino acid sequence                                (SEQ ID NO:55) of nfSP2 is:                                                   TTA GAT GGG CGC ATT GTT GGA GGA CAA GAT GCT GAT ATT GCC AAA                   TAT                                                                            L   D   G   R   I   V   G   G   Q   D   A   D   I   A   K                    Y                                                                              - GGC TAT CAA GCT TCA CTC CAA GTA TTT AAC GAA CAT TTC TGT GGA                GCT                                                                            G   Y   Q   A   S   L   Q   V   F   N   E   H   F   C   G                    A                                                                              - TCA ATA TTG AAT AAT TAT TGG ATT GTC ACA GCA GCT CAT TGC ATA                TAT                                                                            S   I   L   N   N   Y   W   I   V   T   A   A   H   C   I                    Y                                                                              - GAT GAA TTC ACG TAT TCA GTT CGA GTC GGC ACC AGT TTC CAA GGA                AGA                                                                            D   E   F   T   Y   S   V   R   V   G   T   S   F   Q   G                    R                                                                              - CGT GGT TCC GTT CAT CCT GTG GCA CAA ATT ATC AAG CAT CCT GCA                TAC                                                                            R   G   S   V   H   P   V   A   Q   I   I   K   H   P   A                    Y                                                                              - C. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:56), as well as deduced amino acid sequence                                (SEQ ID NO:57) of nfSP4 is:                                                   AGG GAA CAA AAG CTG GAG CTC CAC CGC GGT GCG CCG GCT CTA GAA                   CTA                                                                            R   E   Q   K   L   E   L   H   R   G   A   P   A   L   E                    L                                                                              - GTG GAT CCC CCG GGT CTG CAG GAA TTG GCA CGA GGA TGT TCT TGG                CTG                                                                           V   D   P   P   G   L   Q   E   L   A   R   G   C   S   W                     L                                                                              - TGT TTA GTA GCT ATT CTT TGT GCA GTG GCT GCT GGG CCT ACT AAT                CGC                                                                            C   L   V   A   I   L   C   A   V   A   A   G   P   T   N                    R                                                                              - ATT GTT GGA GGA TTG GAG GCG AAA AAT GGA ATC ACC CCA TTC ATC                GGT                                                                            I   V   G   G   L   E   A   K   N   G   I   T   P   F   I                    G                                                                              - TTC TTT GCA AGC GGA AGA CTA TTT CA                                          F   F   A   S   G   R   L   F                                                 - D. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:58), as well as deduced amino acid sequence                                (SEQ ID NO:59) of nfSP5 is:                                                   ACG AGG TTT CGC TTA GCA ATT GTA TGT GCT CTC GCT GTC TGC ACA                   TTC                                                                           T   R   F   R   L   A   I   V   C   A   L   A   V   C   T   F>                 - GGT GCC AGT GTT CCA GAA CCA TGG AAA AGA TTA GAT GGT AGA ATC                GTA                                                                           G   A   S   V   P   E   P   W   K   R   L   D   G   R   I   V>                 - GGA GGA CAC GAT ACC AGC ATC GAT AAA CAC CCT CAT CAA GTA TCT                TTA                                                                            G   G   H   D   T   S   I   D   K   H   P   H   Q   V   S                    L>                                                                             - TTG TAC TCC AGC CAC AAT TGT GGT GGT TCC TTG ATT GCC AAA AAC                TGG                                                                            L   Y   S   S   H   N   C   G   G   S   L   I   A   K   N                    W>                                                                             - GTT TTG ACT GCA GCT CAT TGC ATT GGA GTT AAC AAA TAC AAT GTC                CGT                                                                            V   L   T   A   A   H   C   I   G   V   N   K   Y   N   V                    R>                                                                             - E. The apparent N-terminal nucleic acid sequence  (SEQ ID                  NO:60), as well as deduced amino acid sequence                                (SEQ ID NO:61) of nfSP6 is:                                                   CCC TCA CTA AAG GGA ACA AAA GCT GGA GCT CCA CCG CGG TGC GCC                   GCT                                                                            P   S   L   K   G   T   K   A   G   A   P   P   R   C   A                    A                                                                              - CTA GAA CTA GTG GAT CCC CCG GGC TGC AGG AAT TCG GCA CGA GCG                TTT                                                                            L   E   L   V   D   P   P   G   C   R   N   S   A   R   A                    F                                                                              - GGT TGG ATT GAG CGC GTC TCA TCT TAC AAG ATA AAG GAT AGA TTA                GAT                                                                            G   W   I   E   R   V   S   S   Y   K   I   K   D   R   L                    D                                                                              - GGG CGC ATT GTT GGA GGA CAA GAT GCT GAT ATT GCC AAA TAT GGC                TAT                                                                            G   R   I   V   G   G   Q   D   A   D   I   A   K   Y   G                    Y                                                                              - CAA GCT TCA CTC CAA GTA CTT AAC GAA CAT TTC TGT GGA GCT                     Q   A   S   L   Q   V   L   N   E   H   F   C   G   A                         - F. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:62), as well as deduced amino acid sequence                                (SEQ ID NO:63) of nfSP7 is:                                                   GCG GTG ATT GTG TCA TTT GTT CTG GCT TGT GCA TTT TCT GTA CAG                   GCT                                                                            A   V   I   V   S   F   V   L   A   C   A   F   S   V   Q                    A                                                                              - CTT CCA TCA AGC AGA ATT GTC AAT GGA CTT GAA GCA GGA GTT GGA                CAA                                                                            L   P   S   S   R   I   V   N   G   L   E   A   G   V   G                    Q                                                                              - TTT CCA ATT CAG GTT TTC TTA GAC TTG ACA AAT ATC AGA GAC GAA                AAA                                                                            F   P   I   Q   V   F   L   D   L   T   N   I   R   D   E                    K                                                                              - TCC AGA TGT GGT GGT GCT TTG TTA TCA GAT TCA TGG GTT TTG ACT                GCT                                                                            S   R   C   G   G   A   L   L   S   D   S   W   V   L   T                    A                                                                              - GCT CAT TGT TTT GAT GAT TTG AAG TCT ATG GTA GTG TCC GTT GGT                GCT                                                                            A   H   C   F   D   D   L   K   S   M   V   V   S   V   G                    A                                                                              - CAT GAT GTC AGC AAA TCT GAA GAA CCT CAC AGG CAA ACC AGG AAA                CCT                                                                            H   D   V   S   K   S   E   E   P   H   R   Q   T   R   K                    P                                                                              - GAA                                                                         E                                                                             - G. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:64), as well as deduced amino acid sequence                                (SEQ ID NO:65) of nfSPl2 is:                                                  GTA CTG ATC GTT TTA GCA GTC ATT GAA TTC GCA TCA GCG TCT TCA                   ATC                                                                            V   L   I   V   L   A   V   I   E   F   A   S   A   S   S                    I                                                                              - GGC TGG AGA ATC GTG GGT GGT GAA AAT GCT AAA GAA AAA TCG GTG                CCC                                                                            G   W   R   I   V   G   G   E   N   A   K   E   K   S   V                    P                                                                              - TAT CAA GTT TCM CTT CGA AAT GCT GAA AAC AAA CAT TTY TGT GGR                GGR                                                                            Y   Q   V   S   L   R   N   A   E   N   K   H   F   C   G                    G                                                                              - H. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:66), as well as deduced amino acid sequence                                (SEQ ID NO:67) of nfSPl3 is:                                                  TTC GGC TTC AAG CTA AGT CAT TTG GTA AGT AAG TAC TGT GCT TGT                   GCA                                                                            F   G   F   K   L   S   H   L   V   S   K   Y   C   A   C                    A                                                                              - TTA GCA TCG GCA CTG AAG TAC TCC ATC GAT CAT GGT CCT CGT ATC                ATC                                                                            L   A   S   A   L   K   Y   S   I   D   H   G   P   R   I                    I                                                                              - GGA GGT GAA GTT GCA GGT GAA GGA TCA GCA CCT TAC CAG GTG TCC                TTA                                                                            G   G   E   V   A   G   E   G   S   A   P   Y   Q   V   S                    L                                                                              - AGA ACC AAG GAA GGA AAT CAT TTT TGC GGT GGA TCA ATA CTA AAT                AAG                                                                            R   T   K   E   G   N   H   F   C   G   G   S   I   L   N                    K                                                                              - CGA TGG GTT GTA ACT GCA GCA CAT TGT CTT GAA CCG GAA ATA TTA                GAT                                                                            R   W   V   V   T   A   A   H   C   L   E   P   E   I   L                    D                                                                              - TCG GTA TAC GTC GGA TCC AAT CAC TTA GAC CGA AAA GGC AGA TAT                TAC                                                                            S   V   Y   V   G   S   N   H   L   D   R   K   G   R   Y                    Y                                                                              - GAC GTA GAA CGG TAT ATA ATT CAT GAA AAA TAT ATA GGA GAA CTA                AAT                                                                            D   V   E   R   Y   I   I   H   E   K   Y   I   G   E   L                    N                                                                              - AAT TTT TAT GCT GAC ATC GGT CTA ATA AAA CTT GAT GGA AGA CTT                AGA                                                                            N   F   Y   A   D   I   G   L   I   K   L   D   G   R   L                    R                                                                              - ATT CAA                                                                     I   Q                                                                         - I. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:68), as well as deduced amino acid sequence                                (SEQ ID NO:69) of nfSPl4 is:                                                  CGG GCT GCA GGA ATT CGG CAC GAG AAG AAA CTG CCA ATA TTA ATC                   GCC                                                                            R   A   A   G   I   R   H   E   K   K   L   P   I   L   I                    A                                                                              - TTG ATC GGA TGC GTT CTT TCT GAA GAA ATA GAG GAT CGC ATT GTC                GGC                                                                            L   I   G   C   V   L   S   E   E   I   E   D   R   I   V                    G                                                                              - GGA ACG GCA GTT GAT ATA AGA GGT TTT CCC TGG CAG GTA TCA ATT                CAA                                                                            G   T   A   V   D   I   R   G   F   P   W   Q   V   S   I                    Q                                                                              - ACC GAA AAC CGT CAT TTT TGT GGT GGT TCT ATT ATC GAT AAA AGC                TGG                                                                            T   E   N   R   H   F   C   G   G   S   I   I   D   K   S                    W                                                                              - ATA TTA ACT GCC GCA CAT TGT GTA CMC GAT ATG AAG ATG TCG AAC                TGG                                                                            I   L   T   A   A   H   C   V   X   D   M   K   M   S   N                    W                                                                              - J. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:70), as well as deduced amino acid sequence                                (SEQ ID NO:71) of nfSPl5 is:                                                  CAC GAG ATT TTA TTA AGC GCA TTA TTT GCA AGT GTA ATT TGC TCC                   TTT                                                                            H   E   I   L   L   S   A   L   F   A   S   V   I   C   S                    F                                                                              - AAC GCG GAA GTA CAA AAT CGA ATC GTT GGT GGC AAT GAT GTA AGT                ATT                                                                            N   A   E   V   Q   N   R   I   V   G   G   N   D   V   S                    I                                                                              - TCA AAA ATT GGG TGG CAA GTA TCT ATT CAA AGT AAT AAA CAA CAT                TTC                                                                            S   K   I   G   W   Q   V   S   I   Q   S   N   K   Q   H                    F                                                                              - TGT GGT GGT TCA ATC ATT GCT AAA GAT GGG TCC                                 C   G   G   S   I   I   A   K   D   G   S                                     - K. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:72), as well as deduced amino acid sequence                                (SEQ ID NO:73) of nfSP16 is:                                                  ATC ATG GCA AAT TTT AGG CTA TTC ACC TTA CTA GCC TTG GTT TCA                   GTA                                                                            I   M   A   N   F   R   L   F   T   L   L   A   L   V   S                    V                                                                              - GCA ACT TCC AAA TAT ATT GAT CCA AGA ATA ATT GGA GGC GAA GAT                GCT                                                                            A   T   S   K   Y   I   D   P   R   I   I   G   G   E   D                    A                                                                              - CCT GAA GGC TCG GCT CCG TAC CAA GTT TCA TTG AGA AAT CAG AAT                CTG                                                                            P   E   G   S   A   P   Y   Q   V   S   L   R   N   Q   N                    L                                                                              - GAG CAT TTC TGT GGT GGT TCC ATT                                             E   H   F   C   G   G   S   I                                                 - L. The apparent N-terminal nucleic acid sequence (SEQ ID                   NO:74), as well as deduced amino acid sequence                                (SEQ ID NO:75) of nfSPl7 is:                                                  GCA CGA GAT CGC ATT GTT GGA GGA TTG GAG GCG AAA AAT GGA TCA                   GCC                                                                            A   R   D   R   I   V   G   G   L   E   A   K   N   G   S                    A                                                                              - CCA TTC ATG GTT TCT TTG CAA GCG GAA GAC TAT TTT CAT TTT TGT                GGA                                                                            P   F   M   V   S   L   Q   A   E   D   Y   F   H   F   C                    G                                                                              - TCC TCT ATT CTG AAT GAG AGA TGG GTT CTT ACT GCT GCT CAC TGT                ATC                                                                            S   S   I   L   N   E   R   W   V   L   T   A   A   H   C                    I                                                                              - CAA CCA AAT GTA CAC AAG TAC GTT TAC GTC GGT TCG AAC AAC GTA                GAA                                                                            Q   P   N   V   H   K   Y   V   Y   V   G   S   N   N   V                    E                                                                              - M. The apparent C-terminal nucleic acid sequence (SEQ ID                   NO:76), as well as deduced amino acid sequence (SEQ ID NO:77)                 of nfSPl2 is:                                                                 CCA ATC CAC GAT AGC CAA TAT GCA CTT TTG CAG ATA TGG GTC AAG                   GGT                                                                            P   I   H   D   S   Q   Y   A   L   L   Q   I   W   V   K                    G>                                                                             - GCA TGT AAG GGT GAT TCC GGT GGC CCC TTA GTC ATC AAT GGA CAA                CTT                                                                            A   C   K   G   D   S   G   G   P   L   V   I   N   G   Q                    L>                                                                             - CAT GGA ATT GTT TCC TGG GGC ATT CCT TGC GCT GTC GCA AGC CTG                ATG                                                                            H   G   I   V   S   W   G   I   P   C   A   V   A   S   L                    M>                                                                             - TAT TCA CAA GAG TTT CTC ATT ATG TCG ATT GGA TTA AAT CCA AAA                TTG                                                                            Y   S   Q   E   F   L   I   M   S   I   G   L   N   P   K                    L>                                                                             - AAT AAA ATT GTT TAG                                                         N   K   I   V   *                                                             - N. The apparent C-terminal nucleic acid sequence (SEQ ID                   NO:78), as well as deduced amino acid sequence                                (SEQ ID NO:79) of nfSPl3 is (the initial GGPL is next                         to the conserved active-site serine):                                         GGA GGT CCT TTG GCA ATC AAT GGT GAA CTT GTT GGT GTT ACT TCA                   TTC                                                                            G   G   P   L   A   I   N   G   E   L   V   G   V   T   S                    F                                                                              - ATT ATG GGG ACA TGT GGA GGA GGA CAT CCT GAT GTC TTC GGT CGA                GTC                                                                            I   M   G   T   C   G   G   G   H   P   D   V   F   G   R                    V                                                                              - CTT GAC TTC AAA CCA TGG ATT GAT TCT CAT ATG GCA AAT GAC GGC                GCT                                                                            L   D   F   K   P   W   I   D   S   H   M   A   N   D   G                    A                                                                              - AAT TCT TTT ATT TAA                                                         N   S   F   I   *                                                          __________________________________________________________________________

Example 12

This Example describes the purification of a flea aminopeptidase of thepresent invention.

The starting material for the isolation of a flea aminopeptidase was aflea midgut lysate preparation that had been depleted of serineproteases by passage over a benzamidine-Sepharose affinity column. Toassay for aminopeptidase activity, the synthetic substrate L-Leucine-AMC(Leu-AMC), which releases a fluorescent AMC leaving group uponproteolytic cleavage, was incubated with the serine protease-depletedflea midgut preparation. Aminopeptidase activity was easily detectablewith as little as 1.2 μg of lysate, both confirming the presence of anaminopeptidase (as indicated in other Examples herein) and allowing forthe detection of aminopeptidase activity in fractions collectedthroughout subsequent fractionation and purification procedures.

Serine protease-depleted flea midgut lysates (samples of about 1.2 μgand about 12 μg) were incubated with Leu-AMC in the presence of thefollowing inhibitors: 1 mM pefabloc, 1 mg/ml trypsin/chymotrypsininhibitor, 1 mg/ml trypsin inhibitor, 1 mM TPCK, 1 μg/ml pepstatin, 10μg/ml E-64, 10 μg/ml of leupeptin, 10 mM EDTA, and 86 μg/ml of bestatin.Only bestatin inhibited the flea protease that cleaved Leu-AMC, whereasboth EDTA and bestatin inhibited the control protease, a leucineaminopeptidase. These results indicated that the flea protease beingcharacterized was an aminopeptidase, but apparently was not ametalloaminopeptidase like the "classic" leucine aminopeptidases.

A flea aminopeptidase was purified using the following protocol. Fleamidgut lysates cleared of serine protease activity were fractionated byanion-exchange chromatography. Those fractions containing aminopeptidaseactivity were pooled and subjected to cation-exchange chromatography,and the resulting fractions were again assayed for activity withL-Leu-AMC in 96-well plates.

Fractions containing aminopeptidase activity were subjected to SDS-PAGEand silver-stained to identify the protein(s) exhibiting that activity.Aminopeptidase activity was found to be associated with proteins thatmigrated at a molecular weight of about 95 kD and about 56 kD whensubjected to SDS-PAGE. The 95 kD and 56 kD proteins may each beaminopeptidases or they may be subunits of a larger enzyme. A number ofknown aminopeptidases are multi-subunit enzymes comprised of subunitsranging from about 45 kD to about 55 kD and from about 90 kD to about 95kD.

Additional purification studies have indicated that the majority ofaminopeptidase activity was found to be associated with the membranepellet preparation and could be solubilized with detergent.Aminopeptidase activity in such preparations was also monitored duringpurification using L-Leu-AMC, and appeared to be associated with the 95kD and 56 kD proteins when active fractions were analyzed by SDS-PAGEand silver staining. The 95 kD and 56 kD protein were co-purified togreater than 90% purity by cation exchange chromatography, affinitychromatography using w-aminohexyl agarose, and C-4 reverse phasechromatography. N-terminal amino acid sequence analysis indicated thatboth isolated aminopeptidases appeared to be blocked at the aminoterminus.

Example 13

This Example describes the isolation of a flea aminopeptidase nucleicacid molecule of the present invention.

A nucleic acid molecule encoding a flea aminopeptidase was isolated inthe following manner. A DNA fragment was PCR amplified from a whole fedflea cDNA expression library (prepared as described in Example 8 usingdegenerate primers, the design of which was based on conserved regionsof bovine lens leucine aminopeptidase (LAP). The specific LAP-basedprimers used included: degenerate LAP sense primer A, corresponding tobovine lens LAP amino acid sequence from about amino acid 247 through257 and having nucleic acid sequence 5' GTW GGW AAA GGW WTW ACW TTY GATTCW GGW GG 3', represented herein as SEQ ID NO:48; and degenerate LAPantisense primer C, corresponding to bovine lens LAP amino acid sequencefrom about amino acid 335 through 329 and having nucleic acid sequence5' CG WCC TTC WGC ATC WGT ATT 3', represented herein as SEQ ID NO:49.Also used were vector primers having SEQ ID NO:13 and SEQ ID NO:14,described in Example 11.

In a first experiment, the LAP primer C having SEQ ID NO:49 and the M13reverse vector primer having SEQ ID NO:13 were used to PCR amplify DNAfragments from the expression library. The resultant PCR products werescreened by hybridization under standard hybridization conditions withLAP primer A having SEQ ID NO:48. A PCR product that hybridized with SEQID NO:48 was subjected to nested (actually semi-nested) PCRamplification using LAP primer C and the T3 vector primer having SEQ IDNO:14. The resulting PCR product, which was about 900 nucleotides inlength (denoted nfAP₉₀₀) and hybridized under standard (i.e., stringent)hybridization conditions with LAP primer A, was cloned into the TA™vector and analyzed by DNA sequence analysis as described in Example 11.

The nucleic acid sequence of a portion of nfAP₉₀₀, namely of nfAP₄₅₃, isrepresented herein as SEQ ID NO:50. Translation of SEQ ID NO:50 yields aprotein of about 151 amino acids, denoted herein as PfAP₁₅₁, the aminoacid sequence of which is represented herein as SEQ ID NO:51. Analysisof SEQ ID NO:51 suggests that the sequence includes a leader segment ofabout 15 amino acids followed by a mature protein that has about 32%identity with the bovine lens LAP. The corresponding bovine and fleanucleotide sequences are about 29% identical.

Example 14

This Example describes the production of an anti-flea midgut proteaseantiserum and its use to inhibit flea protease activity therebysupporting the utility of protease-based vaccines as anti-flea agents.

Anti-flea protease antiserum was produced in the following manner. Arabbit was immunized 3 times with approximately 40-50 μg of a fleamidgut protease preparation that had been affinity-purified usingbenzamidine sepharose as described in Example 7 and then combined withFreund's complete adjuvant for the first immunization and withincomplete adjuvant for the second and third immunizations according tostandard procedures. After the second immunization, endpoint titers ofaround 1:3200 were obtained, while the third immunization boosted theanti-protease titers to about 1:6400. Western blot analysis of theimmunoreactivity of the resultant anti-flea protease antiserum againstthe affinity-purified midgut protease preparation demonstrated thepresence of at least 7-8 reactive protease bands. This was an importantobservation since there are numerous reports in the literature ofdifficulties associated with generating high-titered antisera againstcertain classes of proteases.

To assess the inhibitory activity of the rabbit anti-flea proteaseantiserum against flea midgut proteases, an in vitro assay whichmeasures trypsin activity as a function of absorbance at OD₄₅₀ using adefined protein substrate (i.e., succinylated casein) was establishedusing a commercially available kit (available from Pierce, Rockford,Ill.). In preliminary assays, the proteolytic activity of theaffinity-purified flea midgut protease preparation was about 25-30% ofthe activity observed with the trypsin control. This lower activity isnot unexpected since the flea proteases may require different reactionconditions than the trypsin control for optimal activity. Also, theprimary amino acid sequences determined for the flea proteases asdescribed in Examples 7, 10 and 11 are suggestive of highly specializedfunctions that may require specific substrates for determining optimalactivity. Incubation of the affinity purified midgut proteasepreparation with the succinylated casein substrate in the presence ofabout 500 ng of the rabbit anti-flea protease antibody-containing serumcollected after the second immunization reduced the proteolytic activityof the protease preparation by about 20%. This result, using asuboptimal assay, supports the feasibility of using immunologicalmethods to inhibit flea midgut protease activity.

Using a similar immunization protocol, anti-flea protease antiserum hasalso been generated in cats that has exhibited immunoreactivity, asidentified by Western blot analysis, against several proteases in theaffinity-purified midgut protease preparation. The cat anti-fleaprotease antiserum also reduced proteolytic activity of an affinitypurified midgut protease preparation by about 20%, using the same assayas described for analyzing the rabbit antiserum.

Example 15

This Example demonstrates that flea larvae have predominantlyserine-type proteases.

Newly hatched flea larvae were raised in colony rearing dishes and fedon larval rearing media containing dried bovine blood using standardtechniques. About 300 to 500 larvae were collected at differentdevelopmental stages, homogenized by sonication in flea gut dissectionbuffer (50 mM Tris, 100 mM CaCl₂, pH 8.0) and centrifuged to pellet celldebris. About 25 larval equivalents were incubated with about 2.5 μCi[1,3-³ H]-diisopropylfluorophosphate (DFP) overnight at 4° C. Afterincubation, about 10 larval equivalents were spotted onto filter paper,precipitated with 10% trichloroacetic acid, and counted in a liquidscintillation counter. Reducing SDS-PAGE was performed on samplescomprising about 2.5 larval equivalents, and autoradiography wasperformed using standard techniques. In addition, adult flea midgutproteases were extracted and ³ H-labeled in the same manner and examinedby SDS-PAGE and autoradiography. Analysis of the gel indicated that,based on their ability to be labeled with DFP, larval proteases appearto be predominantly serine-type proteases, the production of whichappears to be induced by blood feeding as occurs in adults. Blood-fed3rd instars appeared to have the highest amount of proteolytic activity.

Example 16

The Example demonstrates that flea feces has proteolytic activity, thatis predominantly due to serine proteases.

Flea feces were collected from fleas fed in flea cages placed on a livecat or in a flea feeding system as described in Example 6 in which thefleas were fed bovine blood. Fresh feces were collected every 9-17hours, resuspended in water at 150 mg feces/ml, and centrifuged topellet insoluble material. The soluble fractions were then assayed usingtwo techniques. Western blot analysis was performed on samples subjectedto reducing SDS-PAGE, each lane having about 40 μg of protein. Blottedproteins were incubated overnight at 4° C. with 1:500 rabbit anti-fleaprotease antiserum produced as described in Example 14. Goat anti-rabbitsecondary antibody was used at 1:2000 to develop the Western blot.Analysis of the Western blot indicated the presence of serine-typeproteases in flea feces. Appearance of such proteases migrating at about25 to about 30 kD in such a system suggests the presence of full-lengthserine proteases.

Zymogram analysis was performed by loading approximately 50 μg proteininto each lane of the electrophoresis gel in non-reducing sample buffer.After electrophoresis, the zymogram gel was soaked in 2.5% Triton-X-100to renature the samples and developed at 37° C. in 50 mM Tris, pH 7.6,200 mM NaCl, 5 mM CaCl₂, 0.02% Brij 35. Coomassie staining the gelrevealed clear plaques where active proteases digested the gelatin inthe gel matrix. Both of these techniques indicated the presence ofserine-type proteases in flea feces.

Example 17

This Example demonstrates that fleas that have fed onantibody-containing blood have antibodies in their feces, suggesting animmunological method to eradicate flea larvae, which feed from fleafeces.

The ability of antibodies in a blood meal to be taken up by fleas, passthrough the midgut and be excreted in the feces was demonstrated in thefollowing manner. A commercially available rabbit antibody againstovalbumin was added at near physiological concentration (i.e., at about2 mg/ml) to the blood meal of adult fleas in a flea feeding system asdescribed in Example 6. Feces were collected at 24 hr and 48 hr afterfeeding and rehydrated in phosphate saline buffer. The rehydrated fecalsamples were subjected to Western blot analysis and shown to containrabbit anti-ovalbumin antibodies that were apparently full-length, usinga rabbit-specific secondary antibody screen against the Fc region ofrabbit antibodies. Supernatants of flea midguts collected at the sametime periods showed residual amounts of rabbit anti-ovalbuminantibodies.

In a second experiment, fleas were fed in a similar manner a blood mealcontaining cat-specific antiserum generated against keyhole limpethemocyanin (KLH) and feces were collected at 24, 48 and 72 hourspost-feeding. The sample collected at 24 hours was divided into halves,with one half rehydrated immediately in PBS while the second half wasrehydrated 7 days later. Fecal samples collected at 48 and 72 hours wereheld for 6 and 5 days, respectively, after collection in desiccated formprior to rehydration. Aliquots of the bloodmeal containing the KLHantiserum fed to the fleas were also sampled at 1 and 2 days. All of therecovered antibodies were reactive against KLH by Western blot analysis,with a pattern or reactivity indistinguishable from the cat anti-KLHserum alone.

These studies demonstrate that antibodies are able to pass through theflea midgut in intact form and are able to maintain theirantigen-binding characteristics, thereby supporting the feasibility ofan immunological method to target larval development, since flea larvaein their normal habitat feed from flea feces.

Example 18

This Example provides additional nucleic acid and deduced amino acidsequences of nucleic acid molecules encoding flea serine proteaseproteins of the present invention, examples of the isolation of whichwere described in Example 11.

A. Flea serine protease clone 1 was determined to comprise flea serineprotease nucleic acid molecule nfSP1₇₇₉, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:80. Translation of SEQ ID NO:80yielded a predicted serine protease protein referred to herein asPfSP1₂₃₂, the amino acid sequence of which is denoted herein as SEQ IDNO:81, SEQ ID NO:80 including an apparent stop codon spanning aboutnucleotides 699 through 701. The N-terminus of the mature form ofPfSP1₂₃₂ apparently occurs at about amino acid 17 of SEQ ID NO:81, and aconserved GWG sequence spans about amino acids 132 through 134 of SEQ IDNO:81. PfSP1₂₃₂ apparently also includes SEQ ID NO:7 (a partial aminoterminal sequence of a purified serine protease protein, the productionof which is described in Example 10). nfSP1₇₇₉ also apparently includesSEQ ID NO:52 and SEQ ID NO:18 and, as such, PfSP1₂₃₂ apparently includesSEQ ID NO:53 and SEQ ID NO:19. A BLAST homology search indicated thatSEQ ID NO:81 was most similar in amino acid sequence to an Anophelesgambiae chymotrypsin II and to a kallikrein. As is the case for any ofthese molecules, variations between sequences may be due to a number offactors, such as but not limited to, sequencing errors or allelicvariation.

B. Flea serine protease clone 2 was determined to comprise flea serineprotease nucleic acid molecule nfSP2₉₄₄, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:82. Translation of SEQ ID NO:82yielded a predicted serine protease protein referred to herein asPfSP2₂₅₅, the amino acid sequence of which is denoted herein as SEQ IDNO:83, SEQ ID NO:82 including an apparent stop codon spanning aboutnucleotides 768 through 770. The N-terminus of the mature form ofPfsP2₂₅₅ apparently occurs at about amino acid 23 of SEQ ID NO:81, and aconserved GWG sequence spans about amino acids 148 through 150 of SEQ IDNO:83. PfSP2₂₅₅ apparently also includes SEQ ID NO:5 (a partial aminoterminal sequence of a purified serine protease protein, the productionof which is described in Example 10). nfSP2₉₄₄ also apparently includesSEQ ID NO:20, and, as such, PfSP2₂₅₅ apparently includes SEQ ID NO:21. ABLAST homology search indicated that SEQ ID NO:83 was most similar inamino acid sequence to a Bombix mori vitellin-degrading proteaseprecursor.

C. Flea serine protease clone 3 was determined to comprise flea serineprotease nucleic acid molecule nfSP3₁₇₇, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:22. Translation of SEQ ID NO:22yielded a predicted serine protease protein referred to herein asPfSP3₅₉, the amino acid sequence of which is denoted herein as SEQ IDNO:23. Flea serine protease protein PfSP3₅₉ includes a conserved GWGsequence that spans about amino acids 1 through 3 of SEQ ID NO:23. ABLAST homology search indicated that SEQ ID NO:23 was most similar inamino acid sequence to a rat trypsinogen.

D. Flea serine protease clone 4 was determined to comprise flea serineprotease nucleic acid molecule nfSP4₆₇₂ described in Example 11. Nucleicacid molecule nfSP4₆₇₂ has nucleic acid sequence SEQ ID NO:16,translation of which yielded a predicted serine protease proteinreferred to herein as PfSP4₂₂₃, the amino acid sequence of which isdenoted herein as SEQ ID NO:17, SEQ ID NO:16 including an apparent stopcodon spanning about nucleotides 670 through 672. As stated above, theN-terminus of the mature form of PfSP4₂₂₃ apparently occurs at aboutamino acid 5 of SEQ ID NO:17, and a conserved GWG sequence spans aboutamino acids 124 through 126 of SEQ ID NO:17. PfSP4₂₂₃ apparentlyincludes SEQ ID NO:41 and a sequence that is very similar to SEQ IDNO:45. nfSP4₆₇₂ also apparently includes SEQ ID NO:24, SEQ ID NO:56(following nucleotide 141 of SEQ ID NO:56) and SEQ ID NO:74. As such,PfSP4₂₂₃ apparently contains SEQ ID NO:25 and SEQ ID NO:75 as well as asequence that is very similar to SEQ ID NO:57 (following amino acid 47of SEQ ID NO:57). A BLAST homology search indicated that SEQ ID NO:17was most similar in amino acid sequence to an A. gambiae chymotrypsin Iprecursor.

E. Flea serine protease clone 5 was determined to comprise flea serineprotease nucleic acid molecules: nfSP5₁₅₇, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:84; and nfSP5₂₁₈, the nucleic acidsequence of which is denoted herein as SEQ ID NO:86. Translation of SEQID NO:84 yielded a predicted serine protease protein referred to hereinas PfSP5₅₂, the amino acid sequence of which is denoted herein as SEQ IDNO:85. The N-terminus of the mature form of the serine protease proteinencoded by flea clone 5 apparently occurs at about amino acid 29 of SEQID NO:85. SEQ ID NO:85 apparently includes the first 10 amino acids ofSEQ ID NO:2 as well as the first 10 amino acids of SEQ ID NO:6. SEQ IDNO:2 and SEQ ID NO:6 are partial N-terminal sequences of purified serineprotease proteins, the production of which are described in Example 10.Translation of SEQ ID NO:86 yielded a predicted serine protease proteinreferred to herein as PfSP5₇₂, the amino acid sequence of which isdenoted herein as SEQ ID NO:87. PfSP5₇₂ includes a conserved GWGsequence spanning about amino acids 14 through 16 of SEQ ID NO:87.nfSP5₂₁₈ apparently includes SEQ ID NO:26, and, as such, PfSP5₇₂apparently includes SEQ ID NO:27. A BLAST homology search indicated thatthe protein encoded by flea clone 5 was most similar in amino acidsequence to a Drosophila trypsin eta precursor.

F. Flea serine protease clone 6 was determined to comprise flea serineprotease nucleic acid molecule nfSP6₉₃₂, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:88. Translation of SEQ ID NO:88yielded a predicted serine protease protein referred to herein asPfSP6₂₅₆, the amino acid sequence of which is denoted herein as SEQ IDNO:89, SEQ ID NO:88 including an apparent stop codon spanningnucleotides 770 through 772. The N-terminus of the mature form ofPfSP6₂₅₆ apparently occurs at about amino acid 26 of SEQ ID NO:89, and aconserved GWG sequence spans about amino acids 149 through 151 of SEQ IDNO:89. PfSP6₂₅₆ apparently also includes SEQ ID NO:4 (a partial aminoterminal sequence of a purified serine protease protein, the productionof which is described in Example 10). nfSP6₉₃₂ also apparently includesSEQ ID NO:28, SEQ ID NO:54 and SEQ ID NO:60 (following about nucleotide111 of SEQ ID NO:60). As such, PfSP6₂₅₆ apparently includes SEQ IDNO:29, SEQ ID NO:55 and SEQ ID NO:61 (following about amino acid 37 ofSEQ ID NO:61). A BLAST homology search indicated that SEQ ID NO:89 wasmost similar in amino acid sequence to a B. mori vitellin-degradingprotease.

G. Flea serine protease clone 7 was determined to comprise flea serineprotease nucleic acid molecule nfSP7₈₉₄, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:90. Translation of SEQ ID NO:90yielded a predicted serine protease protein referred to herein asPfSP7₂₅₅, the amino acid sequence of which is denoted herein as SEQ IDNO:91, SEQ ID NO:90 including an apparent stop codon spanning aboutnucleotides 766 through 768. The N-terminus of the mature form ofPfSP7₂₅₅ apparently occurs at about amino acid 23 of SEQ ID NO:91, and aconserved GWG (in this case GWA) sequence spans about amino acids 152through 154 of SEQ ID NO:91. PfSP7₂₅₅ apparently also includes SEQ IDNO:44. nfSP7₈₉₄ also apparently includes SEQ ID NO:30 and SEQ ID NO:62.As such, PfSP7₂₅₅ apparently includes SEQ ID NO:31 and SEQ ID NO:63. ABLAST homology search indicated that SEQ ID NO:91 was most similar inamino acid sequence to a hornet chymotrypsin II and to collagenase.

H. Flea serine protease clone 8 was determined to comprise flea serineprotease nucleic acid molecule nfSP8₂₉₉, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:92. Translation of SEQ ID NO:92yielded a predicted serine protease protein referred to herein asPfSP8₉₉, the amino acid sequence of which is denoted herein as SEQ IDNO:93. PfSP8₉₉ includes a conserved GWG sequence that spans about aminoacids 31 through 33 of SEQ ID NO:93. nfSP8₂₉₉ also apparently includesSEQ ID NO:32, and, as such, PfSP8₉₉ apparently includes SEQ ID NO:33. ABLAST homology search indicated that SEQ ID NO:93 was most similar inamino acid sequence to a Tachypleus tridentatus coagulation factor B.

I. Flea serine protease clone 9 was determined to comprise flea serineprotease nucleic acid molecule nfSP9₂₆₆, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:94. Translation of SEQ ID NO:94yielded a predicted serine protease protein referred to herein asPfSP9₈₈, the amino acid sequence of which is denoted herein as SEQ IDNO:95. PfSP9₈₈ includes a conserved GWG sequence that spans amino acidsabout 33 through 35 of SEQ ID NO:95. nfSP9₂₆₆ also apparently includesSEQ ID NO:34, and, as such, PfSP9₈₈ apparently includes SEQ ID NO:35. ABLAST homology search indicated that SEQ ID NO:95 was most similar inamino acid sequence to an A. gambiae trypsin 2 precursor.

J. Flea serine protease clone 10 was determined to comprise flea serineprotease nucleic acid molecule nfSP10₃₇₈, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:96. Translation of SEQ ID NO:96yielded a predicted serine protease protein referred to herein asPfSP10₁₂₆, the amino acid sequence of which is denoted herein as SEQ IDNO:97. PfSP10₁₂₆ includes a conserved GWG sequence that spans aboutamino acids 63 through 65 of SEQ ID NO:97. nfSP10₃₇₈ also apparentlyincludes SEQ ID NO:36, and, as such, PfSP10₁₂₆ apparently includes SEQID NO:37. A BLAST homology search indicated that SEQ ID NO:97 was mostsimilar in amino acid sequence to an A. gambiae trypsin 1 precursor.

K. Flea serine protease clone 11 was determined to comprise flea serineprotease nucleic acid molecule nfSP1₂₅₂, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:98. Translation of SEQ ID NO:98yielded a predicted serine protease protein referred to herein asPfSP11₈₄, the amino acid sequence of which is denoted herein as SEQ IDNO:99. PfSP11₈₄ includes a conserved GWG sequence that spans about aminoacids 23 through 25 of SEQ ID NO:99. nfSP11₂₅₂ also apparently includesSEQ ID NO:38, and, as such, PfSP11₈₄ apparently includes SEQ ID NO:39. ABLAST homology search indicated that SEQ ID NO:99 was most similar inamino acid sequence to a Mus musculus plasminogen precursor.

L. Flea serine protease clone 12 was determined to comprise flea serineprotease nucleic acid molecules: nfSP12₁₄₄, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:64; and nfSP12₂₂₅, the nucleic acidsequence of which is denoted herein as SEQ ID NO:100. Translation of SEQID NO:64 yielded a predicted serine protease protein referred to hereinas PfSP12₅₂, the amino acid sequence of which is denoted herein as SEQID NO:65. The N-terminus of the mature form of the serine proteaseprotein encoded by flea clone 12 apparently occurs at about amino acid20 of SEQ ID NO:65. SEQ ID NO:65 apparently includes the first 30 aminoacids of SEQ ID NO:40. Translation of SEQ ID NO:100 yielded a predictedserine protease protein referred to herein as PfSP12₆₉, the amino acidsequence of which is denoted herein as SEQ ID NO:101, SEQ ID NO:100apparently containing a stop codon spanning from about nucleotides 208through 210. nfSP12₂₂₅ apparently includes SEQ ID NO:76, and, as such,PfSP12₆₉ apparently includes SEQ ID NO:77. A BLAST homology searchindicated that the protein encoded by flea clone 12 was most similar inamino acid sequence to an A. gambiae trypsin.

M. Flea serine protease clone 13 was determined to comprise flea serineprotease nucleic acid molecule nfSP13₈₅₀, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:102. Translation of SEQ ID NO:102yielded a predicted serine protease protein referred to herein asPfSP13₂₅₂₁ the amino acid sequence of which is denoted herein as SEQ IDNO:103, SEQ ID NO:102 including an apparent stop codon spanning aboutnucleotides 758 through 760. The N-terminus of the mature form ofPfSP13₂₅₂ apparently occurs at about amino acid 28 of SEQ ID NO:103, anda conserved GWG sequence spans about amino acids 137 through 139 of SEQID NO:103. PfSP13₂₅₂ apparently also includes SEQ ID NO:1 (a partialamino terminal sequence of a purified serine protease protein, theproduction of which is described in Example 10) and SEQ ID NO:42.nfSP13₈₅₀ also apparently includes SEQ ID NO:66 and SEQ ID NO:78 and, assuch, PfSP13₂₅₂ apparently includes SEQ ID NO:67 and SEQ ID NO:79. ABLAST homology search indicated that SEQ ID NO:103 was most similar inamino acid sequence to an A. gambiae chymotrypsin.

N. Flea serine protease clone 14 was determined to comprise flea serineprotease nucleic acid molecule nfSP14₂₁₃, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:68. Translation of SEQ ID NO:68yielded a predicted serine protease protein referred to herein asPfSP14₇₁, the amino acid sequence of which is denoted herein as SEQ IDNO:69. The N-terminus of the mature form of the serine protease proteinencoded by flea clone 12 apparently occurs at about amino acid 29 of SEQID NO:69. SEQ ID NO:69 apparently includes the first 13 amino acids ofSEQ ID NO:43. A BLAST homology search indicated that the protein encodedby SEQ ID NO:69 was most similar in amino acid sequence to an A. gambiaetrypsin.

O. Flea serine protease clone 15 was determined to comprise flea serineprotease nucleic acid molecule nfSP15₂₅₂, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:104. Translation of SEQ ID NO:104yielded a predicted serine protease protein referred to herein asPfSP15₈₄, the amino acid sequence of which is denoted herein as SEQ IDNO:105. The N-terminus of the mature form of the serine protease proteinencoded by flea clone 15 apparently occurs at about amino acid 28 of SEQID NO:105. SEQ ID NO:105 apparently includes SEQ ID NO:46. nfSP15₂₅₂also apparently includes SEQ ID NO:70, and, as such, PfSP15₈₄ apparentlyincludes SEQ ID NO:71. A BLAST homology search indicated that theprotein encoded by SEQ ID NO:105 was most similar in amino acid sequenceto an A. gambiae trypsin.

P. Flea serine protease clone 16 was determined to comprise flea serineprotease nucleic acid molecule nfSP16₁₆₈, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:72. Translation of SEQ ID NO:72yielded a predicted serine protease protein referred to herein asPfSP16₅₆, the amino acid sequence of which is denoted herein as SEQ IDNO:73. The N-terminus of the mature form of the serine protease proteinencoded by flea clone 16 apparently occurs at about amino acid 26 of SEQID NO:73. SEQ ID NO:73 apparently includes SEQ ID NO:47. A BLASThomology search indicated that the protein encoded by SEQ ID NO:73 wasmost similar in amino acid sequence to an acrosin.

Q. Flea serine protease clone 18 was determined to comprise flea serineprotease nucleic acid molecule nfSP18₅₃₄, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:106. Translation of SEQ ID NO:106yielded a predicted serine protease protein referred to herein asPfSP18₁₇₈, the amino acid sequence of which is denoted herein as SEQ IDNO:107. The N-terminus of the mature form of PfSP18₁₇₈ apparently occursat about amino acid 284 of SEQ ID NO:107, and a conserved GWG sequencespans about amino acids 126 through 128 of SEQ ID NO:107. A BLASThomology search indicated that SEQ ID NO:107 was most similar in aminoacid sequence to a chymotrypsin.

R. Flea serine protease clone 19 was determined to comprise flea serineprotease nucleic acid molecule nfSP19₃₅₉, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:108. Translation of SEQ ID NO:108yielded a predicted serine protease protein referred to herein asPfSP19₁₁₉, the amino acid sequence of which is denoted herein as SEQ IDNO:109. A conserved GWG sequence spans about amino acids 69 through 71of SEQ ID NO:109. A BLAST homology search indicated that SEQ ID NO:109was most similar in amino acid sequence to bovine duodenase I.

S. Flea serine protease clone 20 was determined to comprise flea serineprotease nucleic acid molecule nfSP20₈₄₁, the nucleic acid sequence ofwhich is denoted herein as SEQ ID NO:110. Translation of SEQ ID NO:110yielded a predicted serine protease protein referred to herein asPfSP20₂₄₈, the amino acid sequence of which is denoted herein as SEQ IDNO:111, SEQ ID NO:110 including an apparent stop codon spanning aboutnucleotides 746 through 748. The N-terminus of the mature form ofPfSP20₂₄₈ apparently occurs at about amino acid 27 of SEQ ID NO:111, anda conserved GWG sequence spans about amino acids 147 through 149 of SEQID NO:111. PfSP20₂₄₈ apparently also includes SEQ ID NO:2 and SEQ IDNO:6 (partial amino terminal sequences of purified serine proteaseproteins, the production of which is described in Example 10). nfSP20₈₄₁also apparently includes SEQ ID NO:58, and, as such, PfSP20₂₄₈apparently includes SEQ ID NO:59. A BLAST homology search indicated thatSEQ ID NO:111 was most similar in amino acid sequence to a trypsin.

Example 19

This Example provides additional nucleic acid and deduced amino acidsequences of nucleic acid molecules encoding flea aminopeptidaseproteins of the present invention.

The nucleic acid sequence of the remainder of flea aminopeptidasenucleic acid molecule nfAP₉₀₀ was determined and used to design primersto use in combination with a vector primer (i.e., M13 universal primer)to PCR amplify the 3' terminal fragment of the flea aminopeptidasecoding region from a whole fed flea cDNA expression library usingmethods as described in Example 13. The PCR product was subjected to DNAsequencing analysis, and a composite sequence representing a close tofull-length flea aminopeptidase coding region was deduced. The nucleicacid sequence of the composite nucleic acid molecule, referred to hereinas nfAP₁₅₈₀, is denoted herein as SEQ ID NO:112. The primer used toobtain the 3' terminal fragment spans from about nucleotide 849 through877 of SEQ ID NO:112. A probe spanning from about nucleotide 918 through938 of SEQ ID NO:112 was used to verify that the 3' terminal fragmentwas a flea aminopeptidase nucleic acid molecule. The flea aminopeptidasegene-containing sequence of the 3' terminal fragment, referred to hereinas nfAP₇₃₂, spans from about nucleotide 849 through 1580 of SEQ IDNO:112.

Translation of SEQ ID NO:112 yielded a deduced flea aminopeptidaseprotein of about 496 amino acids, denoted herein as PfAP₄₉₆, havingamino acid sequence SEQ ID NO:113. The deduced mature fleaaminopeptidase is about 48% identical to mature bovine leucineaminopeptidase. The corresponding bovine and flea nucleic acid sequencesare about 33% identical.

Example 20

This Example demonstrates the production of certain flea serine proteaseproteins of the present invention.

A. Flea serine protease protein PfSP1₂₁₆ was produced in the followingmanner. An about 670-bp DNA fragment, referred to herein as nfSP1₆₇₀ anddesigned to encode an apparently mature serine protease protein, was PCRamplified from flea serine protease clone 1 using the XhoI-sitecontaining primers F1 sense 5' GAGCTCTCGAGAGTTGTTGGAGGACTGGAAGC 3' (SEQID NO:114) and F1 antisense 5' GGACCTCGAGAATTAGTTATTTTCCATGGTC 3' (SEQID NO:115). The PCR product nfSP1₆₇₀ was digested with XhoI restrictionendonuclease, gel purified and subcloned into expression vector pTrcHisB(available from InVitrogen) that had been digested with XhoI anddephosphorylated. The resultant recombinant molecule, referred to hereinas pHis-nfSP1₆₇₀, was transformed into E. coli HB101 competent cells(available from Gibco BRL, Gaithersburg, Md.) to form recombinant cellE. coli:pHis-nfSP1₆₇₀. The recombinant cell was cultured in enrichedbacterial growth medium containing 0.1 mg/ml ampicillin and 0.1% glucoseat about 32° C. When the cells reached an OD600 of about 0.4-0.5,expression of nfSP1₆₇₀ was induced by the addition of 0.5 mMisopropyl-B-D-thiogalactoside (IPTG), and the cells were cultured forabout 2 hours at about 32° C. Immunoblot analysis of recombinant cell E.coli:pHis-nfSP1₆₇₀ lysates using a T7 tag monoclonal antibody (availablefrom Novagen, Inc., Madison, Wis.) directed against the fusion portionof the recombinant PHIS-PfSP1₂₁₆ fusion protein identified a protein ofthe appropriate size, namely an about 29 kD protein.

B. Flea serine protease protein PfSP2₂₃₃ was produced in the followingmanner. An about 715-bp DNA fragment, referred to herein as nfSP2₇₁₅ anddesigned to encode an apparently mature serine protease protein, was PCRamplified from flea serine protease clone 2 using the XhoI-sitecontaining primers F2 sense 5' GAGCTCTCGAGCATCGTCGGCGGCACCAGTG 3' (SEQID NO:116) and F2 antisense 5' GGACGAATTCTTAAAGACCAGTTTTTTTGCG 3' (SEQID NO:117). The PCR product nfSP2₇₁₅ was digested with XhoI restrictionendonuclease, gel purified and subcloned into expression vector pTrcHisB(available from InVitrogen) that had been digested with XhoI anddephosphorylated. The resultant recombinant molecule, referred to hereinas pHis-nfSP2₇₁₅, was transformed into E. coli HB101 competent cells(available from Gibco BRL) to form recombinant cell E.coli:pHis-nfSP2₇₁₅. The recombinant cell was cultured as described inExample 20A. Immunoblot analysis of recombinant cell E.coli:pHis-nfSP2₇₁₅ lysates using a T7 tag monoclonal antibody (availablefrom Novagen, Inc.) directed against the fusion portion of therecombinant PHIS-PfSP2₂₃₃ fusion protein identified a protein of theappropriate size, namely an about 35-kD protein.

C. Flea serine protease protein PfSP13₂₂₅ was produced in the followingmanner. An about 700-bp DNA fragment, referred to herein as nfSP13₇₀₀and designed to encode an apparently mature serine protease protein, wasPCR amplified from flea serine protease clone 13 using the XhoI-sitecontaining primers F13 sense 5' GAGCTCTCGAGTATCATCGGAGGTGAAGTTGC 3' (SEQID NO:118) and F13 antisense 5' GGACCTCGAGAATTATGCGCCGTCATTTGC 3' (SEQID NO:119). The PCR product nfSP13₇₀₀ was digested with XhoI restrictionendonuclease, gel purified and subcloned into expression vector pTrcHisB(available from InVitrogen) that had been digested with XhoI anddephosphorylated. The resultant recombinant molecule, referred to hereinas pHis-nfSP13₇₀₀, was transformed into E. coli HB101 competent cells(available from Gibco BRL) to form recombinant cell E.coli:pHis-nfSP13₇₀₀. The recombinant cell was cultured as described inExample 20A. Immunoblot analysis of recombinant cell E.coli:pHis-nfSP13₇₀₀ lysates using a T7 tag monoclonal antibody(available from Novagen, Inc.) directed against the fusion portion ofthe recombinant PHIS-PfSP13₂₂₅ fusion protein identified a protein ofthe appropriate size, namely an about 33-kD protein.

Example 21

This Example demonstrates the temporal induction of serine proteases infleas feeding on cats.

Fleas contained in chambers similar to those used for in vitro feedingexperiments were placed on cats and were allowed to feed for variousperiods of time. Upon removal from the cats, soluble extracts of fleamidgut tissues were prepared as described herein. Proteases containedwithin the extracts were quantitated by labeling the extracts with[1,3-³ H] DFP using a method similar to that described in Borovsky etal, 1988, Arch. Insect Biochem. Physiol. 7, 187-210. The labeled sampleseither (a) were precipitated and the radioactivity in the precipitatequantitated or (b) were applied to SDS-PAGE and exposed byautoradiography.

Data generated from counting samples are shown in FIG. 10, which plots[³ H]-DFP in counts per minute (cpm) per 10 flea midguts versus hours offlea feeding on cats. Also shown in FIG. 10 for comparison is a plot ofthe number of eggs laid per female flea per day for chamber-containedfleas feeding on cats for 1, 2, 3 and 4 days, respectively. Theseresults suggest that DFP-labeled proteases (i.e., predominantly serineproteases) are induced in fleas in response to feeding. Induction isquite rapid once feeding begins and, unlike in mosquitos, is sustainedover time. The results also suggest a positive correlation between fleaprotease activity and fecundity.

In order to obtain a profile of the sizes of DFP-labeled proteasestemporally induced in fleas during feeding, samples were applied toSDS-PAGE and autoradiographed. FIG. 11 indicates protein molecularweight standards and samples of soluble flea midgut extracts obtainedfrom fleas having fed for various times on cats (i.e., at, respectively,3, 8, 15, 18, 24, 30, 34, 40, 44, 48, 52, 58, 68, 72, 78 and 88 hours offeeding). Analysis of the results indicates that primarily proteasesmigrating with a molecular weight of about 25-35 kD are induced in aflea relatively soon after the flea has begun feeding (i.e., at leastwithin about 3 to about 8 hours). The amount of such proteases increasesover time for about the first 2 days. Over time, several intenselylabeled bands of lower molecular weight (primarily in the range of about12-15 kD) also appear that may be representative of proteases havingundergone degradation.

Example 22

This example describes the determination of internal amino acid sequenceof a flea aminopeptidase.

About 10,200 cat blood-fed flea guts were dissected into 4 ml GutDissection Buffer (50 mM Tris-HCl, pH 8.0 and 100 mM CaCl₂). Flea gutextracts were prepared by sonicating the flea guts and centrifuging themat about 14,000 rpm for about 20 minutes. The resulting pellet waswashed and briefly sonicated in 2ml Gut Dissection Buffer andcentrifuged again at about 14,000 rpm for about 20 minutes. Theresulting pellet was resuspended and sonicated in 4 ml buffer comprising20 mM NaAc, pH 6.0, 0.1% Brij, complete protease inhibitor cocktail(available from Pierce) and 0.25 mM bestatin; the sonicate wascentrifuged at about 14,000 rpm for about 20 minutes. Both the pelletand supernatant were recovered. The pellet was re-sonicated andcentrifuged as above, and the resulting supernatant was combined withthe original supernatant.

The pooled supernatant was applied to a polyCAT cation exchange HPLCcolumn and protein was eluted with a NaCl gradient ranging from 0M to 1MNaCl in 20 mM NaAc, pH 6.0. Fractions collected from the column wereassayed by H-Leu-AMC fluorescence, and active fractions were pooled andapplied to a C-1 reverse phase HPLC column (TMS 250, Toso Hass).Proteins were eluted from the column using an acetonitrile gradient in0.1% TFA in water, the gradient ranging between 20% and 100%acetonitrile. Proteins contained in fractions from the column wereanalyzed by SDS-PAGE gel electrophoresis and silver staining. Theresults of the gel electrophoresis indicated the presence of an about 95kDa protein in some of the fractions. This protein correlates with theabout 95 kDa protein described in Example 12 which was identified usingmembrane pellet from flea midgut lysates.

To determine internal amino acid sequence of the 95 kDa protein, thosefractions containing the 95 kDa protein were pooled, dried and digestedwith BNPS-Skatole for about 72 hours at room temperature. TheBNPS-Skatole digest was separated by 18% Tris-glycine PAGE gelelectrophoresis and blotted onto PVDF membrane. A major band of about 28kDa was cut out and N-terminally sequenced using techniques as describedin Example 7. A partial N-terminal amino acid sequence of the internalpeptide was obtained, namely LATTQFQATHARSAFPCFDEPAM (denoted herein SEQID NO:167).

Example 23

This example describes the cloning and sequencing of another fleaaminopeptidase nucleic acid molecule.

Primer APN3 corresponding to a conserved region in Manduca sexta and rataminopeptidases, having nucleic acid sequence 5' CCC AAA TTT TCC ATW GCNCCN GC 3' (N indicating any nucleotide; represented herein as SEQ IDNO:166) was used in combination with primer M13 Reverse primer (SEQ IDNO:13) to PCR amplify a portion of a flea aminopeptidase gene from abovine blood-fed whole flea cDNA expression library as described abovein Example 8. The resulting product of the PCR amplification was dilutedabout 1:50 and used as a template in a second, semi-nested PCRamplification using a primer APN3 in combination with degenerate primerAPN1C, designed using SEQ ID NO:167 (described in Example 22), havingnucleic acid sequence 5' CAA TTY CAA GCT ACY CAT GC 3' (representedherein as SEQ ID NO:168). The resulting PCR product, named nfAP2₃₈₃, wasapproximately 383-bp when visualized on a 1% agarose gel. The PCRproduct nfAP2₃₈₃ was gel purified and cloned into the TA Vector® System,and subjected to standard DNA sequencing techniques. The nucleotidesequence of nfAP2₃₈₃ is denoted SEQ ID NO:169. Translation of SEQ IDNO:169 yielded a deduced flea aminopeptidase protein of about 127 aminoacids, denoted herein as PfAP2₁₂₇, having amino acid sequence SEQ IDNO:170.

The PCR product nfAP2₃₈₃ was labelled with ³² p and used as a probe toscreen a bovine blood-fed whole flea phage expression library usingstandard hybridization techniques. A single plaque purified clone wasisolated, which included a 2100-nucleotide insert, referred to herein asnfAP2₂₁₀₀. Partial nucleic acid sequence was obtained using standardtechniques from the 5' end of nfAP2₂₁₀₀, to yield a flea aminopeptidasenucleic acid molecule named nfAP2₅₃₇ having nucleic acid sequence SEQ IDNO:171. Translation of SEQ ID NO:171 suggests that nucleic acid moleculenfAP2₅₃₇ encodes a non-full-length flea aminopeptidase protein of about178 amino acids, referred to herein as PfAP₁₇₈, having amino acidsequence SEQ ID NO:172, assuming the first codon spans from aboutnucleotide 2 through about nucleotide 4 of SEQ ID NO:171. SEQ ID NO:172contains SEQ ID NO:167.

Flea aminopeptidase nucleic acid sequence SEQ ID NO:171 was comparedwith additional nucleic acid sequences characterized from otherorganisms. The nucleic acid sequence is about 50% identical to Manducasexta aminopeptidase N nucleotides between corresponding regions of thetwo nucleic acid molecules.

Example 24

This example describes the cloning and sequencing of a flea cysteineprotease nucleic acid molecule.

A flea cysteine protease nucleic acid molecule, referred to herein asnfCP1₅₇₃ was produced by PCR amplification using the following method.Primer Cal3F (designed to obtain a calreticulin gene), having nucleicacid sequence 5' TTG GGA TAC ACT TTG ACT GTT AAC C 3', representedherein as SEQ ID NO:173 was used in combination with the M13 universalprimer, to PCR amplify, using standard techniques, a DNA fragment from abovine blood-fed whole flea CDNA expression library as described abovein Example 8. Surprisingly, the isolated DNA fragment correlated with acysteine protease nucleic acid sequence. Sequence from this DNA fragmentwas used to design primer Cys1R, having the nucleic acid sequence 5' GTGAGC AAC CAT TAT TTC CAT ATC 3', represented herein as SEQ ID NO:174,which was used in a second PCR amplification in combination with the M13reverse primer. A third PCR amplification was performed using primerCys1F, having the nucleic acid sequence 5' CTT TCC TCA CAA TAC CAC CAAGGA AGC 3', represented herein as SEQ ID NO:176, in combination with theM13 universal primer. A fourth PCR amplification was performed usingprimer Cys2F, having the nucleic acid sequence 5' CTT GTA CGA TTG TCTCAA CAG GC 3', represented herein as SEQ ID NO:175, in combination withthe M13 universal primer. The resulting PCR products were each gelpurified and cloned into the TA Vector® System, and subjected tostandard DNA sequencing techniques. A composite nucleic acid sequencerepresenting a flea cysteine protease coding region was deduced,referred to herein as nfCP1₅₇₃, was deduced and is denoted herein as SEQID NO:177. Translation of SEQ ID NO:177 suggests that nucleic acidmolecule nfCP1₅₇₃ encodes a non-full-length flea cysteine proteaseprotein of about 191 amino acids, referred to herein as PfCP1₁₉₁, havingamino acid sequence SEQ ID NO:178, assuming the first codon spans fromabout nucleotide 1 through about nucleotide 3 of SEQ ID NO:177.

The nucleic acid and amino acid sequences of the nfCP1₅₇₃ nucleic acidmolecule and PfCP1₁₉₁ protein, respectively, were compared to knownnucleic acid and amino acid sequences using a Genbank homology search.SEQ ID NO:178 was found to be similar to the amino acid sequence of P.sativum cysteine protease. The most highly conserved region ofcontinuous similarity between SEQ ID NO:178 and P. sativum cysteineprotease amino acid sequences spans from about amino acid 71 throughabout amino acid 165 of SEQ ID NO:178 and from about amino acid 17through about amino acid 168 of the P. sativum cysteine protease, therebeing about 42% identity between the two regions. Comparison of thenucleic acid sequence encoding amino acids from about 205 through about492 of nfCP1₅₇₃ indicate that those regions are about 54% identical.

Example 25

This example describes the cloning and sequencing of certain flea serineprotease nucleic acid molecules.

Additional serine protease cDNA nucleic acid molecules have beenisolated in a manner similar to that described in Example 8. The actualprimers used in PCR amplification of serine protease nucleic acidmolecules from a bovine blood-fed flea cDNA expression library (producedas described in Example 8) included cat-try #2 (SEQ ID NO:12) incombination with either M13 reverse primer (SEQ ID NO:13) or H57 primer(SEQ ID NO:15). The resultant PCR products were gel purified and clonedinto the TA Vector™. Two recombinant TA vector clones were isolated andfound to correspond to previously cloned serine protease genes. Thesenewly cloned nucleic acid molecules were subjected to nucleic acidsequencing using the Sanger dideoxy chain termination method, asdescribed in Sambrook et al., ibid.

A. A nucleic acid sequence of the flea serine protease nucleic moleculecorresponding to flea clone 5 (produced using primers cat try #2 and M13reverse), namely nfSP5₈₀₆ is represented herein as SEQ ID NO:120. SEQ IDNO:84 and SEQ ID NO:86 are both contained within the sequence of thenucleic acid molecule nfSP5₈₀₆ Translation of SEQ ID NO:120 suggeststhat nucleic acid molecule nfSP5₈₀₆ encodes a close to full-length fleaserine protease protein of about 245 amino acids, referred to herein asPfSP5₂₄₅, having amino acid sequence SEQ ID NO:121, assuming an openreading frame in which the first codon spans from about nucleotide 2through about nucleotide 4 of SEQ ID NO:120 and a stop codon spanningfrom about nucleotide 737 through about nucleotide 739 of SEQ ID NO:120.A Genbank homology search revealed most homology between SEQ ID NO:120and a Gallus gallus trypsin gene, there being about 52% identity betweencorresponding regions of the two nucleic acid molecules.

B. A nucleic acid sequence of the flea serine protease nucleic moleculecorresponding to flea clone 11 (produced using primers cat try #2 andM13 reverse), namely nfSP11₃₀₇, is represented herein as SEQ ID NO:124.SEQ ID NO:38 and SEQ ID NO:98 are within the sequence of the nucleicacid molecule nfSP11₃₀₇ Translation of SEQ ID NO:124 suggests thatnucleic acid molecule nfSP11₃₀₇ encodes a non-full-length flea serineprotease protein of about 102 amino acids, referred to herein asPfSP11₁₀₂, having amino acid sequence SEQ ID NO:125, assuming the firstcodon spans from about nucleotide 1 through about nucleotide 3 of SEQ IDNO:124.

C. A nucleic acid sequence of the flea serine protease nucleic moleculecorresponding to flea clone 39 (produced using primers cat try #2 andH57), namely nfSP39₂₆₇, is represented herein as SEQ ID NO:164.Translation of SEQ ID NO:164 suggests that nucleic acid moleculenfSP39₂₆₇ encodes a non-full-length flea serine protease protein ofabout 90 amino acids, referred to herein as PfSP39₈₉, having amino acidsequence SEQ ID NO:165, assuming the first codon spans from aboutnucleotide 1 through about nucleotide 3 of SEQ ID NO:164.

Example 26

This example describes the cloning and sequencing of certain flea serineprotease nucleic acid molecules.

A. Bovine Blood-Fed Library

Certain flea serine protease cDNA nucleic acid molecules have beenisolated in a manner similar to that described in Example 8, using twonucleic acid molecules as probes to screen a bovine blood-fed flea cDNAexpression library (produced as described in Example 8), cat-try #1 (SEQID NO:11) and cat-try #2 (SEQ ID NO:12). Two clones that hybridizedstrongly to the probes were isolated and subjected to nucleic acidsequencing using the Sanger dideoxy chain termination method, asdescribed in Sambrook et al., ibid.

1. The nucleic acid sequence of a flea serine protease nucleic moleculecorrelating to flea clone 8, namely nfSP8₄₃₆ is represented herein asSEQ ID NO:122. SEQ ID NO:92 is within the sequence of the nucleic acidmolecule nfSP8₄₃₆ Translation of SEQ ID NO:122 yields a protein of about145 amino acids, denoted PfSP8₁₄₅, having amino acid sequence SEQ IDNO:123, assuming the first codon spans from about nucleotide 2 throughabout nucleotide 4 of SEQ ID NO:122. A Genbank homology search revealedmost homology between SEQ ID NO:122 and an Anopheles gambiae trypsinprecursor gene, there being about 48% identity between correspondingregions of the two nucleic acid molecules.

2. The nucleic acid sequence of a flea serine protease nucleic moleculecorresponding to flea clone 12, namely nfSP12₇₅₈ is represented hereinas SEQ ID NO:126. SEQ ID NO:64 and SEQ ID NO:100 are both containedwithin the sequence of the nucleic acid molecule nfSP12₇₅₈. Translationof SEQ ID NO:126 yields a protein of about 246 amino acids, denotedPfSP12₂₄₆, having amino acid sequence SEQ ID NO:127, assuming an openreading frame in which the first codon spans from about nucleotide 1through about nucleotide 3 of SEQ ID NO:126 and a stop codon spanningfrom about nucleotide 739 through about nucleotide 741 of SEQ ID NO:127.A Genbank homology search revealed most homology between SEQ ID NO:126and a rat trypsinogen gene, there being about 57% identity betweencorresponding regions of the two nucleic acid molecules.

B. Cat Blood-Fed Library

Certain flea serine protease cDNA genes have been isolated from a catblood-fed flea cDNA expression library by screening the library with thecat-try #1 (SEQ ID NO:11) and cat-try #2 (SEQ ID NO:12) probes. The catblood-fed flea library was produced in a similar manner as the bovineblood-fed flea library (described in Example 8) except the fleas werefed on cat blood. Two clones that hybridized strongly to the probes wereisolated and subjected to nucleic acid sequencing using methodsdescribed above.

1. The nucleic acid sequence of one of the flea serine protease nucleicmolecules, namely nfSP26₆₁₀ is represented herein as SEQ ID NO:140.Translation of SEQ ID NO:140 yields a non-full-length sequence of about185 amino acids, denoted PfSP26₁₈₅, having amino acid sequence SEQ IDNO:141, assuming the first codon spans from about nucleotide 1 throughabout nucleotide 3 of SEQ ID NO:140. A Genbank homology search revealedmost homology between SEQ ID NO:141 and a Aedes aegypti trypsin proteinsequence, there being about 48% identity between corresponding regionsof the two amino acid sequences.

2. The nucleic acid sequence of a flea serine protease nucleic molecule,namely nfSP27₃₈₆ is represented herein as SEQ ID NO:142. Translation ofSEQ ID NO:142 yields a protein of about 128 amino acids, denotedPfSP27₁₂₈, having amino acid sequence SEQ ID NO:143, assuming the firstcodon spans from about nucleotide 1 through about nucleotide 3 of SEQ IDNO:142.

Example 27

This example describes the cloning and sequencing of certain flea serineprotease nucleic acid molecules.

Certain serine protease cDNA nucleic acid molecules have been isolatedfrom reverse transcriptase PCR amplification of mRNA isolated from catblood-fed whole fleas. The mRNA was isolated from fleas gathered over 72hours after the initiation of feeding on cat blood. As such, the mRNAcomprised a mixture of mRNA isolated at different time points over 72hours. The mRNA was isolated using ground-up fleas, extracting totalflea RNA using Tri-Reagent (available from Molecular Research Center,Cincinnati, Ohio)and an Invitrogen Fast Track™ RNA isolation kit(available from Invitrogen, Inc. San Diego, Calif.). cDNA wassynthesized using a Stratagene RT-PCR kit (available from Stratagene,Inc, San Diego, Calif.). Primers used for first-strand cDNA synthesisincluded an equal molar mixture of the following: 5' dT-2VT3' and 5'dT-2VC3' (as provided in a differential display kit, available fromOperon Technologies, Inc. Alameda, Calif.).

The actual primers used in the PCR amplification of the cDNA describedabove included cat-try #2 (SEQ ID NO:12) used in combination with H57primer (SEQ ID NO:15). The resultant PCR products were gel purified andcloned into the TA Vectors. Six recombinant TA vector clones wereisolated and the nucleic acid molecules were subjected to nucleic acidsequencing using analysis as described above.

A. A nucleic acid sequence of one of the flea serine protease nucleicmolecules, namely nfSP23₄₂₃, is represented herein as SEQ ID NO:134.Translation of SEQ ID NO:134 suggests that nucleic acid moleculenfSP23₄₂₃ encodes a non-full-length flea serine protease protein ofabout 141 amino acids, referred to herein as PfSP23₁₄₁, having aminoacid sequence SEQ ID NO:135, assuming the first codon spans from aboutnucleotide 1 through about nucleotide 3 of SEQ ID NO:134. A Genbankhomology search revealed most homology between SEQ ID NO:134 and a Homosapiens plasminogen precursor gene, there being about 51% identitybetween corresponding regions of the two nucleic acid molecules.

B. Another nucleic acid sequence of a flea serine protease nucleicmolecule, namely nfSP24₄₁₀, is represented herein as SEQ ID NO:136.Translation of SEQ ID NO:136 suggests that nucleic acid moleculenfSP24₄₁₀ encodes a non-full-length flea serine protease protein ofabout 136 amino acids, referred to herein as PfSP24₁₃₆, having aminoacid sequence SEQ ID NO:137, assuming the first codon spans from aboutnucleotide 1 through about nucleotide 3 of SEQ ID NO:136. A Genbankhomology search revealed most homology between SEQ ID NO:137 and anAnopheles gambiae chymotrypsin protein sequence, there being about 38%identity between corresponding regions of the two amino acid sequences.

C. Another nucleic acid sequence of a flea serine protease nucleicmolecule, namely nfSP33₄₂₆, is represented herein as SEQ ID NO:154.Translation of SEQ ID NO:154 suggests that nucleic acid moleculenfSP33₄₂₆ encodes a non-full-length flea serine protease protein ofabout 142 amino acids, referred to herein as PfSP33₁₄₂, having aminoacid sequence SEQ ID NO:155, assuming the first codon spans from aboutnucleotide 1 through about nucleotide 3 of SEQ ID NO:154. A Genbankhomology search revealed most homology between SEQ ID NO:155 and aDrosophila serine protease stubble protein sequence, there being about45% identity between corresponding regions of the two amino acidsequences.

D. Another nucleic acid sequence of one of the flea serine proteasenucleic molecule, namely nfSP36₁₉₇, is represented herein as SEQ IDNO:158. SEQ ID NO:158 represents a partial sequence of a PCR amplifiednucleic acid molecule nfSP36₅₀₀. Translation of SEQ ID NO:158 suggeststhat nucleic acid molecule nfSP36₁₉₇ encodes a non-full-length fleaserine protease protein of about 65 amino acids, referred to herein asPfSP36₆₅, having amino acid sequence SEQ ID NO:159, assuming the firstcodon spans from about nucleotide 1 through about nucleotide 3 of SEQ IDNO:158. A Genbank homology search revealed most homology between SEQ IDNO:159 and a Drosophila melanogaster easter protein sequence, therebeing about 42% identity between corresponding regions of the two aminoacid sequences.

E. Another nucleic acid sequence of a flea serine protease nucleicmolecule, namely nfSP38₃₄₁, is represented herein as SEQ ID NO:162.Translation of SEQ ID NO:162 suggests that nucleic acid moleculenfSP38₃₄₁ encodes a non-full-length flea serine protease protein ofabout 113 amino acids, referred to herein as PfSP38₁₁₃, having aminoacid sequence SEQ ID NO:163, assuming the first codon spans from aboutnucleotide 3 through about nucleotide 5 of SEQ ID NO:162. A Genbankhomology search revealed most homology between SEQ ID NO:163 and a rattrypsinogen protein sequence, there being about 30% identity betweencorresponding regions of the two amino acid sequences.

F. A nucleic acid sequence of one of the flea serine protease nucleicmolecules, namely nfSP34₃₉₀, is represented herein as SEQ ID NO:156.Translation of SEQ ID NO:156 suggests that nucleic acid moleculenfSP4₃₉₀ encodes a non-full-length flea serine protease protein of about130 amino acids, referred to herein as PfSP34₁₃₀, having amino acidsequence SEQ ID NO:157, assuming the first codon spans from aboutnucleotide 1 through about nucleotide 3 of SEQ ID NO:156. A Genbankhomology search revealed most homology between SEQ ID NO:157 and aDrosophila melanogaster Delta precursor protein sequence, there beingabout 33% identity between corresponding regions of the two amino acidsequences.

Example 28

This example describes the cloning and sequencing of a flea serineprotease nucleic acid molecule.

A serine protease cDNA nucleic acid molecule was isolated in a mannersimilar to that described in Example 8. The actual primers used in PCRamplification of the serine protease nucleic acid molecule from a catblood-fed whole flea cDNA expression library (produced as described inExample 26) included cat-try #2 (SEQ ID NO:12) in combination with M13reverse primer (SEQ ID NO:13). The resulting PCR product was diluted1:25 and used as a template in a second PCR reaction using the forwardvector primer T3 (SEQ ID NO:15) in combination with the reverse primer(derived from the nucleic acid sequence of nfSP33₇₇₈, described inExample 27) having the nucleic acid sequence 5' ATT CCT CGT GGT TCA GTCGCT C 3', represented herein as SEQ ID NO:188. The resultant PCR productwas gel purified and cloned into the TA Vector™. The clones weresubjected to nucleic acid sequencing as described above.

A nucleic acid sequence of a flea serine protease nucleic molecule,namely nfSP33₇₇₈ is represented herein as SEQ ID NO:189. As expected,SEQ ID NO:189 includes a portion of SEQ ID NO:154 Translation of SEQ IDNO:189 suggests that nucleic acid molecule nfSP33₇₇₈ encodes anon-full-length flea serine protease protein of about 259 amino acids,referred to herein as PfSP33₂₅₉, having amino acid sequence SEQ IDNO:190, assuming the first codon spans from about nucleotide 2 throughabout nucleotide 4 of SEQ ID NO:189. A Genbank homology search revealedmost homology between SEQ ID NO:189 and a Drosophila serine proteasestubble gene, there being about 54% identity between nucleotides 23-778of SEQ ID NO:189 and nucleotides 2324-3064 of the Drosophila serineprotease stubble gene.

Example 29

This example describes the cloning and sequencing of another flea serineprotease nucleic acid molecule.

Using the method described in Example 26, a cDNA clone of a flea serineprotease was obtained using mRNA isolated from bovine blood-fed wholefleas. The resulting cDNA was used as a template in PCR amplificationusing the primers cat-try #2 (SEQ ID NO:12) used in combination with H57primer (SEQ ID NO:15). The resultant PCR products were gel purified andcloned into the TA Vector™. One recombinant TA vector clone was isolatedand the flea serine protease nucleic acid molecule and denoted nFS37₅₀₀was subjected to nucleic acid sequencing as described in Sambrook etal., ibid.

The nucleic acid sequence of part of the flea serine protease nucleicmolecule nFS37₅₀₀, namely nfSP37₂₆₁, is represented herein as SEQ IDNO:160. Translation of SEQ ID NO:160 suggests that nucleic acid moleculenfSP37₂₆₁ encodes a non-full-length sequence of a flea serine proteaseprotein of about 87 amino acids, referred to herein as PfSP37₈₇, havingamino acid sequence SEQ ID NO:161, assuming the first codon spans fromabout nucleotide 1 through about nucleotide 3 of SEQ ID NO:160. AGenbank homology search revealed most homology between SEQ ID NO:161 anda chicken trypsinogen protein sequence, there being about 31% identitybetween corresponding regions of the two amino acid sequences.

Example 30

This example describes the cloning and sequencing of certain larval fleaserine protease nucleic acid molecules.

Certain serine protease cDNA nucleic acid molecules have been isolatedfrom a mixed instar larval CDNA library produced using 1st, 2nd and 3rdinstar larvae fed on cat blood, by PCR amplification. The actual primersused in the PCR amplification included either cat-try #2 (SEQ ID NO:12)in combination with either H57 primer (SEQ ID NO:15)or M13 reverseprimer (SEQ ID NO:13). The resultant PCR products were gel purified andcloned into the TA Vector™. Three recombinant TA vector clones wereisolated containing PCR products using cat-try #2 and M13 reverse asprimers and one clone was isolated containing PCR products using cat-try#2 and H57 primers. These newly cloned nucleic acid molecules weresubjected to nucleic acid sequencing as described above.

A. A nucleic acid sequence of one of the larval flea serine proteasenucleic molecules isolated using cat-try #2 and M13 reverse primers,namely nfSP29₆₁₂ is represented herein as SEQ ID NO:146. Translation ofSEQ ID NO:146 suggests that nucleic acid molecule nfSP29₆₁₂ encodes aclose to full-length flea serine protease protein of about 204 aminoacids, referred to herein as PfSP29₂₀₄, having amino acid sequence SEQID NO:147, assuming an open reading frame in which the first codon spansfrom about nucleotide 10 through about nucleotide 12 of SEQ ID NO:146. AGenbank homology search revealed most homology between SEQ ID NO:146 anda rat trypsinogen gene, there being about 50% identity betweencorresponding regions of the two nucleic acid molecules.

B. Another nucleic acid sequence of one of the larval flea serineprotease nucleic molecules isolated using cat-try #2 and M13 reverseprimers, namely nfSP30₆₄₁, is represented herein as SEQ ID NO:148.Translation of SEQ ID NO:148 suggests that nucleic acid moleculenfSP30₆₄₁ encodes a non-full-length flea serine protease protein ofabout 213 amino acids, referred to herein as PfSP30 ₂₁₃, having aminoacid sequence SEQ ID NO:149, assuming the first codon spans from aboutnucleotide 3 through about nucleotide 5 of SEQ ID NO:148. A Genbankhomology search revealed most homology between SEQ ID NO:148 and aAnopheles gambiae trypsin gene, there being about 52% identity betweencorresponding regions of the two nucleic acid molecules.

C. Another nucleic acid sequence of one of the larval flea serineprotease nucleic molecules isolated using cat-try #2 and M13 reverseprimers, namely nfSP31₆₂₆, is represented herein as SEQ ID NO:150.Translation of SEQ ID NO:150 suggests that nucleic acid moleculenfSP31₆₂₆ encodes a non-full-length flea serine protease protein ofabout 208 amino acids, referred to herein as PfSP⁻ 208, having aminoacid sequence SEQ ID NO:151, a assuming the first residue spans fromabout nucleotide 3 through about nucleotide 5 or from a putative startcodon spanning from about nucleotide 6 to about nucleotide 8 of SEQ IDNO:150. A Genbank homology search revealed homology between SEQ IDNO:150 and an Anopheles gambiae trypsin gene, there being about 52%identity between corresponding regions of the two nucleic acidmolecules.

D. Another nucleic acid sequence of one of the larval flea serineprotease nucleic molecules isolated using cat-try #2 and H57 primers,namely nfSP32₄₃₃, is represented herein as SEQ ID NO:152. Translation ofSEQ ID NO:152 suggests that nucleic acid molecule nfSP32₄₃₃ encodes anon-full-length flea serine protease protein of about 144 amino acids,referred to herein as PfSP32₁₄₄, having amino acid sequence SEQ IDNO:153, assuming the first codon spans from about nucleotide 1 throughabout nucleotide 3 of SEQ ID NO:152. A Genbank homology search revealedmost homology between SEQ ID NO:152 and an Anopheles gambiae trypsingene, there being about 52% identity between corresponding regions ofthe two nucleic acid molecules.

Example 31

This example describes the cloning and sequencing of another flea serineprotease nucleic acid molecule.

A bovine blood-fed whole flea cDNA library (prepared as described inExample 8) was immunoscreened with antiserum collected from a rabbitthat was immunized with a collection of flea salivary gland productsreferred to as fspN. Immunoscreening was performed as follows. NewZealand White rabbit antiserum developed against fspN flea salivaproducts was used in the immunoscreening protocols described in thepicoBlue™ Immunoscreening Kit instruction manual, available fromStratagene, Inc. The methods for preparation of the CDNA expressionlibraries for immunoscreening, i.e., expression of the cDNA clones andprocedures for transferring lambda phage plaques to membranes forimmunoscreening, are described in the ZAP-CDNA Synthesis Kit instructionmanual, also available from Stratagene, Inc., La Jolla, Calif.

A nucleotide sequence for a flea serine protease nucleic acid moleculenamed nfsp15₈₁₅ is denoted as SEQ ID NO:128 and corresponds to SEQ IDNO:70. Translation of SEQ ID NO:128 suggests that nucleic acid moleculenfsp15₈₁₅ encodes a close to full-length flea serine protease protein ofabout 254 amino acids, referred to herein as Pfsp15₂₅₄, having aminoacid sequence SEQ ID NO:129, assuming an open reading frame in which thefirst codon spans from about nucleotide 1 through about nucleotide 3 ofSEQ ID NO:128 and a stop codon spanning from about nucleotide 763through about nucleotide 765 of SEQ ID NO:128. A Genbank homology searchrevealed homology between SEQ ID NO:128 and an Anopheles gambiae trypsingene, there being about 52% identity between corresponding regions ofthe two nucleic acid molecules.

Example 32

This example describes the cloning and sequencing of additional fleaserine protease nucleic acid molecules.

Certain flea serine protease cDNA nucleic acid molecules have beenisolated in a manner similar to that described in Example 8, using twonucleic acid molecules as probes to screen an unfed flea cDNA expressionlibrary, nfSP8₂₉₉ (SEQ ID NO:92) and nfSP19₃₅₉ (SEQ ID NO:108). A clonethat hybridized strongly to the probes was isolated and subjected tonucleic acid sequencing as described above.

A. The nucleic acid sequence of the flea serine protease nucleicmolecule, namely nfSP19₈₅₅, is represented herein as SEQ ID NO:130. SEQID NO:108 is within the sequence of the nucleic acid molecule nfSP19₈₅₅.Translation of SEQ ID NO:130 yields an apparent full-length protein ofabout 253 amino acids, denoted PfSP19₂₅₃, having amino acid sequence SEQID NO:131, assuming the first codon, an apparent start codon, spans fromabout nucleotide 1 through about nucleotide 3 of SEQ ID NO:130. AGenbank homology search revealed most homology between SEQ ID NO:130 andan Aedes aegypti trypsin, there being about 53% identity betweencorresponding regions of both nucleic acid molecules.

B. The nucleic acid sequence of another flea serine protease nucleicmolecule, namely nfSP25₈₆₄, is represented herein as SEQ ID NO:138.Translation of SEQ ID NO:138 yields a protein of about 260 amino acids,denoted PfSP25₂₆₀, having amino acid sequence SEQ ID NO:139, assumingthe first codon spans from about nucleotide 2 through about nucleotide 4of SEQ ID NO:138 and a stop codon spanning from about nucleotide 782through about nucleotide 784 of SEQ ID NO:139. A Genbank homology searchrevealed most homology between SEQ ID NO:139 and an Anopheles gambiaechymotrypsin protein sequence, there being about 34% identity betweencorresponding regions of the two amino acid sequences.

Example 33

This example describes the cloning and sequencing of another flea serineprotease nucleic acid molecule.

A flea serine protease cDNA nucleic acid molecule. has been isolated ina manner similar to that described in Example 8, using nfSP11₂₅₂ (SEQ IDNO:98) as a probe to screen an bovine blood-fed flea cDNA expressionlibrary (produced as described in Example 8). A clone that hybridizedstrongly to the probe was isolated and subjected to nucleic acidsequencing using the Sanger dideoxy chain termination method, asdescribed in Sambrook et al., ibid.

The nucleic acid sequence of the flea serine protease nucleic molecule,namely nfSP21₅₉₅, is represented herein as SEQ ID NO:132. Translation ofSEQ ID NO:132 yields a protein of about 198 amino acids, denotedPfSP21₁₉₈, having amino acid sequence SEQ ID NO:133, assuming the firstcodon spans from about nucleotide 2 through about nucleotide 4 of SEQ IDNO:132 and a putative stop codon spanning from about nucleotide 596 toabout nucleotide 598. A Genbank homology search revealed most homologybetween SEQ ID NO:133 and Tachypleus tridentatus coagulation factor Gprotein sequence, there being about 45% identity between correspondingregions of the two amino acid sequences.

Example 34

This example describes the isolation and characterization of a 31 kDflea serine protease.

Guts from about 1500 fleas that had been fed on cat blood for about 24hours were dissected in Gut Dissection Buffer (50 mM Tris 8.0, 100 mMCaCl₂). The guts were disrupted by freezing and thawing 4 times,followed by sonication. The resulting extracts were clarified bycentrifugation for 20 minutes at 14,000 rpm in a microfuge at 4° C. Thesupernatant was recovered.

The gut supernatant was loaded onto a 3 ml column comprisingp-aminobenzamidine cross-linked to Sepharose beads (Sigma), previouslyequilibrated in Benzamidine Column Buffer (50 mM Tris 8.0, 100 mM CaCl₂,400 mM NaCl). The supernatant was incubated on the column for about 10min. Unbound protein was slowly washed off the column using BenzamidineColumn Buffer until no protein was detectable by Bradford Assay (BioRad).

Proteases bound to the benzamidine column were then eluted using 10 mlBenzamidine Column Buffer supplemented with 10 mM p-aminobenzamidine(brought to pH 8.0 with NaOH). Proteases in the eluant were concentratedand diafiltered into a volume of about 0.3 ml Gut Dissection Bufferusing a Microcon 3 concentrator (Amicon).

About 120 μl of the concentrated eluant was further concentrated to avolume of about 30 μl. Proteases contained in this concentrate wereresolved by gel electrophoresis on a 14% Tris-Glycine electrophoresisgel (15 μl per lane=approximately 300 gut equivalents per lane). Afterelectrophoresis, the separated proteases were blotted onto a PVDFmembrane using a CAPS buffer (10 mM CAPS pH 11, 0.5 mM DTT) The membranewas stained with Coomassie Brilliant Blue. A dominant protein band ofabout 31 kDa was visualized. The membrane was then used for automatedN-terminal sequencing (described in Example 7). A partial N-terminalamino acid sequence of the flea protease was determined to beIVGGEDVDISTCGWC (denoted SEQ ID NO:179).

Example 35

This example describes the isolation and characterization of a 31 kDflea serine protease contained in a formulation having IgGase activity(i.e., ability to proteolyze immunoglobulin G proteins).

Cat blood-fed flea gut extracts were prepared and selected on abenzamidine column as described above in Example 32. IgG proteaseactivity was assayed by incubating at 37° C., overnight, the benzamidineeluent with cat immunoglobulin G proteins (IgG) purified on Protein Asepharose. The ability of the flea gut benzamidine eluant to digest catIgG was detected by resolving the samples by gel electrophoresis througha 14% SDS-PAGE gel and silver staining the gel using standard methods.The marked decrease (compared with control samples lacking proteaseactivity) of a 50 kDa band on the silver stained gel, representing catIgG heavy chain, indicated that the benzamidine eluant contains IgGprotease activity.

The benzamidine eluant was then purified on a PolyPropylaspartamidehydrophobic interaction chromatography (HIC) column by applying theeluant to the column in buffer containing 0.1 M KPO₄, pH 6.5 and 2 M(NH₄)₂ SO₄. Proteases bound to the column were eluted using an ammoniumsulfate gradient of 2 M to 0 M in HIC column buffer. Column fractionswere tested for IgG protease activity using the method described above.Fractions containing IgG protease activity were pooled and applied to aPolyCat cation exchange column in 20 M sodium acetate, pH 6. Theproteins were eluted using a sodium chloride gradient of 0 M to 1 M NaClin 20 M sodium acetate. Fractions eluted from the column were tested forIgG protease activity and then each fraction was resolved byelectrophoresis using SDS-PAGE. Fractions having the highest levels ofIgG protease activity included a protein band that migrated at about 31kDa on the SDS-PAGE gel. Weaker protease activity corresponded to anabout 28 kDa band.

The 31 kDa protein present on the SDS-PAGE gel was used for N-terminalamino acid sequencing using the blotting method described above. Apartial N-terminal amino acid sequence was determined to beIVGGEDVDIST(C) GWQI(S) FQ(S) ENLHF(C) GG(S) IIAPK (denoted herein as SEQID NO:184). A comparison of SEQ ID NO:184 and SEQ ID NO:179 (describedin Example 34) indicates a single residue difference between the twoamino acid sequences at residue 15 of each sequence (i.e., Q and V,respectively). Since SEQ ID NO:184 correlates with IgGase activity, thedata suggests that the larval protein containing SEQ ID NO:179 hasIgGase activity.

Example 36

This example describes the cloning and sequencing of a 31 kDa fleaserine protease contained in a formulation having IgGase activity.

A flea protease nucleic acid molecule was isolated from a cat blood-fedwhole flea library (described in Example 23) and a bovine blood-fedwhole flea library (described in Example 8) by PCR amplification. Theactual primers used in the PCR amplification included FP31A primerdesigned using the N-terminal amino acid sequence SEQ ID NO:179, theprimer having the nucleic acid sequence 5' GAA GAT GTW GAT ATT TCW ACATGT GG 3' (SEQ ID NO:185) used in combination with the M13 universalprimer. The resultant PCR products were gel purified and cloned into theTA Vector™ and subjected to nucleic acid sequencing as described above.

A FP31B primer (5' GAA AAT GAA ATC CAC TTA AAC ATT ACG 3', (representedherein as SEQ ID NO:186) was designed using the DNA sequence of a DNAfragment from a bovine blood-fed cDNA library. A flea protease cDNAnucleic acid molecule was isolated by PCR amplification of the catblood-fed whole flea library and the bovine blood-fed whole flea librarydescribed above by PCR amplification. PCR amplification was performedusing the FP31B primer in combination with M13 reverse primer. Theresulting PCR products were then diluted 1:25, and used as a templatefor a second PCR reaction using primer FP31C, having the sequence 5' CTCTTA TTG TAC GAG GGA TGC 3' (denoted herein SEQ ID NO:187) in combinationwith T3 primer (SEQ ID NO:14). The resulting nested PCR product wascloned into TA Vector™ and subjected to DNA sequencing.

The nucleic acid sequence of the resulting flea serine protease nucleicmolecule, namely nfSP28₉₂₃ is represented herein as SEQ ID NO:144.Translation of SEQ ID NO:144 yields a protein of about 267 amino acids,denoted PfSP28₂₆₇, having amino acid sequence SEQ ID NO:145, assuming anopen reading frame in which the putative start codon spans from aboutnucleotide 8 through about nucleotide 10 of SEQ ID NO:144 or from aboutnucleotide 11 through about nucleotide 13, and a stop codon spanningfrom about nucleotide 803 through about nucleotide 805 of SEQ ID NO:144.SEQ ID NO:145 contains SEQ ID NO:179 except Q is substituted for C, andSEQ ID NO:184 A Genbank homology search revealed most homology betweenSEQ ID NO:144 and Bombix mori vitellin-degrading protease gene, therebeing about 53% identity between corresponding regions of the twonucleic acid sequences.

Example 37

This example describes ³ H-DFP labelling of larval serine proteases.

About 100 unfed larvae, 100 1st instar larvae, and 100 3rd Instar larvaewere collected in 100 μl Gut Dissection Buffer (50 mM Tris 8.0, 100 mMCaCl₂). About 400 μl of water was added to the collected larvae, whichwere then sonicated. The sonicates were clarified by centrifugation at15,000 RPM in an SS-34 rotor for 30 minutes at 4° C. The supernatantfrom each larval sonicate was recovered and concentrated to a volume ofabout 120 μl (1.2 μl per larval equivalent).

Samples containing about 25 larval equivalents (about 30 μl) werelabeled with about 2.5 μCi of 3H-diisopropylfluorophosphate (DFP;obtained from New England Nuclear) and incubated at 4° C. for 18 hours.Following the incubation period, 5 larval equivalents of each larvalstage (about 6 μl) were run on a 14% Tris Glycine SDS-PAGE gel. The gelwas then soaked in Entensify (obtained from New England Nuclear) toenhance the tritium signal for autoradiography, and dried. The dried gelwas then exposed to X-ray film (Kodak XO-mat) for about 3 days at -70°C.

The results indicate that gut extracts from unfed larvae do not containdetectable amounts of serine proteases (FIG. 12, lane A), while both fed1st instar larvae (FIG. 12, lane B) and fed 3rd instar larvae (FIG. 12,lane C) produce serine proteases. In particular, fed 1st instar larvaeprimarily produce a serine protease having a molecular weight of about25 kD; and fed 3rd instar larvae produce about serine proteases havingmolecular weights of about 25 kD, 28 kD and 31 kD. The approximate sizeof standard molecular weight protein markers are shown in FIG. 12.

Example 38

This example describes the determination of partial N-terminal aminoacid sequences for several larval serine proteases.

About 10,300 3rd Instar larvae were collected in Gut Dissection Buffer(50 mM Tris, pH 8.0, 100 mM CaCl₂). The larvae were homogenized bysonication and clarified by centrifugation at 15,000 rpm in an SS-34rotor for 30 min., at 4° C. The supernatant was recovered. The 3rdInstar supernatant was mixed with 5 ml of p-aminobenzamidinecross-linked to Sepharose beads (Sigma) equilibrated in BenzamidineColumn Buffer (50 mM Tris pH 8.0, 100 mM CaCl₂, 400 mM NaCl). Thesupernatant was rocked with the beads overnight at 4° C. The beads werewashed in about 45 ml Benzamidine Column Buffer to remove unboundprotein. The beads were then mixed 2 hours at 4° C. with about 10 ml ofBenzamidine Column Buffer containing 100 mM p-aminobenzamidine (pH 8.0adjusted with NaOH) to elute proteins bound to the beads. The elutedproteins were then collected. The elution process was repeated oncemore. The eluted protein was concentrated by ultrafiltration with aCentriprep 10 concentrator (Amicon). The concentrate was diluted withGut Dissection Buffer to a final volume of about 5 ml.

Partial N-terminal amino acid sequence of proteins eluted from the beadswas obtained using the method described in Example 32. Two proteinshaving molecular weights of about 25 kDa and about 26 kDa wereidentified on the Coomassie Brilliant Blue stained membranes. PartialN-terminal amino acid sequence obtained for the protein having amolecular weight of about 25 kDa is IVGGVSVNINDYGYQLSLQSNGR, denotedherein as SEQ ID NO:181. Partial N-terminal amino acid sequence obtainedfor the protein having a molecular weight of about 26 kDa isIVGGHDTSIKQHPYQV, denoted herein as SEQ ID NO:180.

Example 39

This Example demonstrates the production of certain flea serine proteaseproteins of the present invention.

A. Flea serine protease protein PfSP1₂₁₆ was in the following manner.Flea serine protease nucleic acid molecule nfSP1₆₇₀, produced asdescribed in Example 20, was digested with XhoI restrictionendonuclease, gel purified and subcloned into expression vector λP_(R)/T² ori/S10HIS-RSET-A9 (the production of which is described in Tripp etal, International PCT Publication No. WO 95/24198, published Sep. 14,1995; see, in particular, Example 7), that had been digested with XhoIand dephosphorylated. The resultant recombinant molecule, referred toherein as pHisCro-nfSP1₆₇₀, was transformed into E. coli HB101 competentcells (available from Gibco BRL) to form recombinant cell E. coli:pHisCro-nfSP1₆₇₀. The recombinant cell was cultured as described inExample 20. Flea serine protease protein PfSP1₂₁₆ was purified by nickelchelation chromatography followed by reverse phase high performanceliquid chromatography (HPLC). Immunoblot analysis of the purifiedPfSP1₂₁₆ indicated that rabbit anti-flea protease antiserum, produced asdescribed in example 14, selectively bound to PfSP1₂₁₆.

B. Flea serine protease protein PfSP2₂₃₃ was produced in the followingmanner. Flea serine protease nucleic acid molecule nfSP2₇₁₅, produced asdescribed in Example 20, was digested with XhoI restrictionendonuclease, gel purified and subcloned into expression vector λP_(R)/T² ori/S10HIS-RSET-A9 as described in Example 39A. The resultantrecombinant molecule, referred to herein as pHisCro-nfSP2₇₁₅ wastransformed into E. coli HB101 competent cells (available from GibcoBRL) to form recombinant cell E. coli:pHisCro-nfSP2₇₁₅. The recombinantcell was cultured as described in Example 20. Flea serine proteaseprotein PfSP2₂₃₃ was purified by nickel chelation chromatographyfollowed by reverse phase HPLC. Immunoblot analysis of the purifiedPfSP2₂₃₃ indicated that rabbit anti-flea protease antiserum, produced asdescribed in example 14, selectively bound to PfSP2₂₃₃.

C. Flea serine protease protein PfSP13₂₂₅ was produced in the followingmanner. Flea serine protease nucleic acid molecule nfSP13₇₀₀, producedas described in Example 20, was digested with XhoI restrictionendonuclease, gel purified and subcloned into expression vector λP_(R)/T² ori/S10HIS-RSET-A9 as described in Example 39A. The resultantrecombinant molecule, referred to herein as pHisCro-nfSP13₇₀₀, wastransformed into E. coli HB101 competent cells (available from GibcoBRL) to form recombinant cell E. coli:pHisCro-nfSP13₇₀₀. The recombinantcell was cultured as described in Example 20. Flea serine proteaseprotein PfSP13₂₂₅ was purified by nickel chelation chromatographyfollowed by reverse phase HPLC. Immunoblot analysis of the purifiedPfSP13₂₂₅ indicated that rabbit anti-flea protease antiserum, producedas described in example 14, selectively bound to PfSP13₂₂₅.

D. Flea serine protease protein PfSP20₂₂₂ was produced in the followingmanner. An about 669-bp DNA fragment, referred to herein as nfSP20₆₆₉,and designed to encode an apparently mature serine protease protein, wasPCR amplified from flea serine protease clone 20 using the XhoI-sitecontaining primer F27-S (sense) 5' GAG CTC TCG AGA ATC GTA GGA GGA CACGAT AC 3' (SEQ ID NO:183) and the EcoRI-site containing primer F20-A(antisense) 5' G GAC GAA TTC TTA AAC ACC AGA CAC TTC CTT G 3' (SEQ IDNO:182). The PCR product nfSP20₆₆₉ was digested with XhoI and EcoRIrestriction endonucleases, gel purified and subcloned into expressionvector λP_(R) /T² ori/S10HIS-RSET-A9 as described in Example 39A. Theresultant recombinant molecule, referred to herein as pHisCro-nfSP20₆₆₉,was transformed into E. coli HB101 competent cells (available from GibcoBRL) to form recombinant cell E. coli:pHisCro-nfSP20₆₆₉. The recombinantcell was cultured as described in Example 20. Immunoblot analysis ofrecombinant cell E. coli:pHisCro-nfSP20₆₆₉ lysates using a T7 tagmonoclonal antibody (available from Novagen, Inc.) directed against thefusion portion of the recombinant PHISCRO-PfSP20₂₂₂ fusion proteinidentified a protein of the appropriate size, namely an about 31-kDprotein. Flea serine protease protein PfSP20₂₂₂ was purified by nickelchelation chromatography followed by reverse phase HPLC. Immunoblotanalysis of the purified PfSP20₂₂₂ indicated that rabbit anti-fleaprotease antiserum, produced as described in example 14, selectivelybound to PfSP20 ₂₂₂.

Example 40

This example describes that various flea serine protease nucleic acidmolecules described in the foregoing examples can be obtained frommultiple sources.

Nucleic acid molecules corresponding to flea clone 4 have been obtainedfrom a bovine blood-fed whole flea library (described in Example 8), acat blood-fed whole flea library (described in Example 26), an unfedwhole flea library (described in Example 8), and a mixed instar wholeflea library (described in Example 30). Nucleic acid moleculescorresponding to flea clone 5 have been obtained from a bovine blood-fedwhole flea library and a cat blood-fed whole flea library. Nucleic acidmolecules corresponding to flea clone 6 have been obtained from a bovineblood-fed whole flea library, a cat blood-fed whole flea library and anunfed whole flea library. Nucleic acid molecule corresponding to fleaclone 7 have been obtained from a bovine blood-fed whole flea library,and a cat blood-fed whole flea library. Nucleic acid moleculescorresponding to flea clone 8 have been obtained from a bovine blood-fedwhole flea library and an unfed whole flea library. Nucleic acidmolecules corresponding to flea clone 12 have been obtained from abovine blood-fed whole flea library and a cat blood-fed whole flealibrary. Nucleic acid molecules corresponding to flea clone 13 have beenobtained from a bovine blood-fed whole flea library, a cat blood-fedwhole flea library, and an unfed whole flea library. Nucleic acidmolecules corresponding to flea clone 20 have been obtained from abovine blood-fed whole flea library, a cat blood-fed whole flea library,and an unfed whole flea library. Nucleic acid molecules corresponding toflea clone 28 have been obtained from a bovine blood-fed whole flealibrary and a cat blood-fed whole flea library.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                   - -  - - (1) GENERAL INFORMATION:                                             - -    (iii) NUMBER OF SEQUENCES: 190                                         - -  - - (2) INFORMATION FOR SEQ ID NO:1:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 32 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                               - - Ile Ile Gly Gly Glu Val Ala Gly Glu Gly Se - #r Ala Pro Tyr Gln        Val                                                                             1               5   - #                10  - #                15              - - Ser Leu Arg Thr Lys Glu Gly Asn His Phe Se - #r Gly Gly Ser Ile Leu                  20      - #            25      - #            30                   - -  - - (2) INFORMATION FOR SEQ ID NO:2:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                               - - Xaa Val Gly Gly His Asp Thr Ser Ile Asp Xa - #a His Pro His Gln Val      1               5   - #                10  - #                15               - - Thr                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:3:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                               - - Ile Val Gly Gly Ala Asp Ala Ala Pro Gly As - #n Ala Pro Phe Gln Val      1               5   - #                10  - #                15               - - Ser Leu Arg Asp Lys Gly                                                              20                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:4:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 34 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                               - - Ile Val Gly Gly Gln Asp Ala Asp Ile Ala Ly - #s Tyr Gly Tyr Gln Ala      1               5   - #                10  - #                15               - - Ser Leu Gln Val Phe Asn Glu His Phe Xaa Gl - #y Ala Xaa Ile Leu Asn                  20      - #            25      - #            30                   - - Asn Tyr                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:5:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 25 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                               - - Ile Val Gly Gly Thr Asp Val Asn Ile Glu As - #n Phe Gly Trp Gln Val      1               5   - #                10  - #                15               - - Ser Leu Phe Asp Arg Asn Gly His Phe                                                  20      - #            25                                          - -  - - (2) INFORMATION FOR SEQ ID NO:6:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                               - - Ile Val Gly Gly His Asp Thr Ser Ile Asp Ly - #s His Pro Phe Gln Val      1               5   - #                10  - #                15               - - Ser Leu Ile Asp Lys Asn                                                              20                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:7:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                               - - Val Val Gly Gly Leu Glu Ala Ala Glu Gly Se - #r Ala Pro Tyr Gln Val      1               5   - #                10  - #                15               - - Xaa Leu Gln Trp Gly Asn Phe                                                          20                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:8:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                               - - Ile Val Gly Gly Glu Asp Ala Glu Leu Gly Gl - #u Xaa Pro Thr Gln          1               5   - #                10  - #                15               - -  - - (2) INFORMATION FOR SEQ ID NO:9:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                               - - Asp Glu Asp Gly Lys Asp Asp Ser Ala Pro Gl - #y Glu Ile                  1               5   - #                10                                      - -  - - (2) INFORMATION FOR SEQ ID NO:10:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino - #acids                                                  (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (ix) FEATURE:                                                                  (A) NAME/KEY: Protein                                                         (B) LOCATION: 1..9                                                            (D) OTHER INFORMATION: - #/note= "At pos. aa 2, Xaa = Gln or                       Asn"                                                            - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                              - - Cys Xaa Gly Asp Ser Gly Gly Pro Leu                                      1               5                                                              - -  - - (2) INFORMATION FOR SEQ ID NO:11:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                              - - TAAWGGWCCW CCYGAATCTC CCTGGCA          - #                  - #                 27                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:12:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                              - - TAAWGGWCCA GARTCTCCTT GACA          - #                  - #                    24                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:13:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 19 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                              - - GGAAACAGCT ATGACCATG             - #                  - #                      - # 19                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:14:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                              - - ATTAACCCTC ACTAAAG             - #                  - #                      - #   17                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:15:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "probe"                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                              - - TGGGTWGTWA CWGCWGCWCA TTG           - #                  - #                    23                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:16:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 672 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..672                                                          (D) OTHER INFORMATION: - #/note= "At pos. aa 224, substitute                       Xaa."                                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                              - - GCA CGA GAT CGC ATT GTT GGA GGA TTG GAG GC - #G AAA AAT GGA TCA GCC           48                                                                       Ala Arg Asp Arg Ile Val Gly Gly Leu Glu Al - #a Lys Asn Gly Ser Ala             1               5 - #                 10 - #                 15              - - CCA TTC ATG GTT TCT TTG CAA GCG GAA GAC TA - #T TTT CAT TTT TGT GGA           96                                                                       Pro Phe Met Val Ser Leu Gln Ala Glu Asp Ty - #r Phe His Phe Cys Gly                        20     - #             25     - #             30                  - - TCC TCT ATT CTG AAT GAG AGA TGG GTT CTT AC - #T GCT GCT CAC TGT ATC          144                                                                       Ser Ser Ile Leu Asn Glu Arg Trp Val Leu Th - #r Ala Ala His Cys Ile                    35         - #         40         - #         45                      - - CAA CCA AAT GTA CAC AAG TAC GTT TAC GTC GG - #T TCG AAC AAC GTA GAA          192                                                                       Gln Pro Asn Val His Lys Tyr Val Tyr Val Gl - #y Ser Asn Asn Val Glu                50             - #     55             - #     60                          - - GTA GGC GGA ACA CAC TAC GAA ATC GAA AAA GC - #T TTC TAT CAC GAA GAA          240                                                                       Val Gly Gly Thr His Tyr Glu Ile Glu Lys Al - #a Phe Tyr His Glu Glu            65                 - # 70                 - # 75                 - # 80       - - TAT GAT GGA GTA GAT CTT GTA GAT CAT GAT GT - #G ATT GAT CAA AGT GAG          288                                                                       Tyr Asp Gly Val Asp Leu Val Asp His Asp Va - #l Ile Asp Gln Ser Glu                            85 - #                 90 - #                 95              - - ACA AAC ATT GAT TTA ATG AAG TGT CAA CCC AT - #T AAA TTA CGA AGA AAG          336                                                                       Thr Asn Ile Asp Leu Met Lys Cys Gln Pro Il - #e Lys Leu Arg Arg Lys                       100      - #           105      - #           110                  - - CCA CTC GTT GGT GGT GAG GAA TTG AGA GCA GT - #A GGC TGG GGA AAT ACA          384                                                                       Pro Leu Val Gly Gly Glu Glu Leu Arg Ala Va - #l Gly Trp Gly Asn Thr                   115          - #       120          - #       125                      - - AAT TCA GCA GGG GAA AAT TTT CCA TTG AAA CT - #T CAA GAA TTG TAC GTG          432                                                                       Asn Ser Ala Gly Glu Asn Phe Pro Leu Lys Le - #u Gln Glu Leu Tyr Val               130              - #   135              - #   140                          - - AAA GCT TTG ACT AAT GAG GAG TGC AAA GCT AA - #A TCA CCA ATT CCA CCA          480                                                                       Lys Ala Leu Thr Asn Glu Glu Cys Lys Ala Ly - #s Ser Pro Ile Pro Pro           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - ACG ACC CAA GTC TGC ACA CTT TTG GAA AAG AA - #T CAT GGT GTA TGC        TCG      528                                                                    Thr Thr Gln Val Cys Thr Leu Leu Glu Lys As - #n His Gly Val Cys Ser                          165  - #               170  - #               175              - - GGA GAT TCT GGT GGT CCA TTG CTT TTG GAT GG - #C GAG CAA GTT GGC ATT          576                                                                       Gly Asp Ser Gly Gly Pro Leu Leu Leu Asp Gl - #y Glu Gln Val Gly Ile                       180      - #           185      - #           190                  - - GCC TCA TTT GTT ATC TTC AAA TGC GCA ATG GG - #G TAC CCT GAC TAT TTC          624                                                                       Ala Ser Phe Val Ile Phe Lys Cys Ala Met Gl - #y Tyr Pro Asp Tyr Phe                   195          - #       200          - #       205                      - - ACA AGA TTG TCT CTA TAT GTA GAT TGG ATT GA - #A CAA CAC ATG GAT TAA          672                                                                       Thr Arg Leu Ser Leu Tyr Val Asp Trp Ile Gl - #u Gln His Met Asp Xaa               210              - #   215              - #   220                          - -  - - (2) INFORMATION FOR SEQ ID NO:17:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 224 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                              - - Ala Arg Asp Arg Ile Val Gly Gly Leu Glu Al - #a Lys Asn Gly Ser Ala        1               5 - #                 10 - #                 15              - - Pro Phe Met Val Ser Leu Gln Ala Glu Asp Ty - #r Phe His Phe Cys Gly                   20     - #             25     - #             30                  - - Ser Ser Ile Leu Asn Glu Arg Trp Val Leu Th - #r Ala Ala His Cys Ile               35         - #         40         - #         45                      - - Gln Pro Asn Val His Lys Tyr Val Tyr Val Gl - #y Ser Asn Asn Val Glu           50             - #     55             - #     60                          - - Val Gly Gly Thr His Tyr Glu Ile Glu Lys Al - #a Phe Tyr His Glu Glu       65                 - # 70                 - # 75                 - # 80       - - Tyr Asp Gly Val Asp Leu Val Asp His Asp Va - #l Ile Asp Gln Ser Glu                       85 - #                 90 - #                 95              - - Thr Asn Ile Asp Leu Met Lys Cys Gln Pro Il - #e Lys Leu Arg Arg Lys                  100      - #           105      - #           110                  - - Pro Leu Val Gly Gly Glu Glu Leu Arg Ala Va - #l Gly Trp Gly Asn Thr              115          - #       120          - #       125                      - - Asn Ser Ala Gly Glu Asn Phe Pro Leu Lys Le - #u Gln Glu Leu Tyr Val          130              - #   135              - #   140                          - - Lys Ala Leu Thr Asn Glu Glu Cys Lys Ala Ly - #s Ser Pro Ile Pro Pro      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Thr Thr Gln Val Cys Thr Leu Leu Glu Lys As - #n His Gly Val Cys        Ser                                                                                             165  - #               170  - #               175             - - Gly Asp Ser Gly Gly Pro Leu Leu Leu Asp Gl - #y Glu Gln Val Gly Ile                  180      - #           185      - #           190                  - - Ala Ser Phe Val Ile Phe Lys Cys Ala Met Gl - #y Tyr Pro Asp Tyr Phe              195          - #       200          - #       205                      - - Thr Arg Leu Ser Leu Tyr Val Asp Trp Ile Gl - #u Gln His Met Asp Xaa          210              - #   215              - #   220                          - -  - - (2) INFORMATION FOR SEQ ID NO:18:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 156 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..156                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                              - - GGG TGG GGA AGA CTT GGA GCT AAC TTG AAT GG - #A CCG AAT GAA CTC CAA           48                                                                       Gly Trp Gly Arg Leu Gly Ala Asn Leu Asn Gl - #y Pro Asn Glu Leu Gln             1               5 - #                 10 - #                 15              - - GAA CTT AAC ACT GTC ACA TTA AGC CAC CAG CA - #A TGT GTA AGA CAA CAA           96                                                                       Glu Leu Asn Thr Val Thr Leu Ser His Gln Gl - #n Cys Val Arg Gln Gln                        20     - #             25     - #             30                  - - ATT TAT CCA GTA TAC GAC AGC CAA CTT TGC AC - #A TTT GTT GGC AGT GGA          144                                                                       Ile Tyr Pro Val Tyr Asp Ser Gln Leu Cys Th - #r Phe Val Gly Ser Gly                    35         - #         40         - #         45                      - - CGA GGC GCC TGC            - #                  - #                      - #      156                                                                  Arg Gly Ala Cys                                                                    50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:19:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 52 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                              - - Gly Trp Gly Arg Leu Gly Ala Asn Leu Asn Gl - #y Pro Asn Glu Leu Gln        1               5 - #                 10 - #                 15              - - Glu Leu Asn Thr Val Thr Leu Ser His Gln Gl - #n Cys Val Arg Gln Gln                   20     - #             25     - #             30                  - - Ile Tyr Pro Val Tyr Asp Ser Gln Leu Cys Th - #r Phe Val Gly Ser Gly               35         - #         40         - #         45                      - - Arg Gly Ala Cys                                                               50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:20:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 168 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..168                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                              - - GGA TGG GGC AAA TTA AGT GAA TCA GGA CCC AA - #G CCA GTA AAT CTA CAA           48                                                                       Gly Trp Gly Lys Leu Ser Glu Ser Gly Pro Ly - #s Pro Val Asn Leu Gln             1               5 - #                 10 - #                 15              - - GGA GTA AAA GTG CCT TAT GTG ACC AAG ATA CA - #T GCT CTG ACA GCT ACG           96                                                                       Gly Val Lys Val Pro Tyr Val Thr Lys Ile Hi - #s Ala Leu Thr Ala Thr                        20     - #             25     - #             30                  - - TCT TTG CAG GTA AAA GAT ATC ACC GAA AAC AT - #G TTG TGT GCC GGA GTT          144                                                                       Ser Leu Gln Val Lys Asp Ile Thr Glu Asn Me - #t Leu Cys Ala Gly Val                    35         - #         40         - #         45                      - - AGA AGA GGT GGC AAG GAC TCC TGC     - #                  - #                   168                                                                     Arg Arg Gly Gly Lys Asp Ser Cys                                                    50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:21:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 56 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                              - - Gly Trp Gly Lys Leu Ser Glu Ser Gly Pro Ly - #s Pro Val Asn Leu Gln        1               5 - #                 10 - #                 15              - - Gly Val Lys Val Pro Tyr Val Thr Lys Ile Hi - #s Ala Leu Thr Ala Thr                   20     - #             25     - #             30                  - - Ser Leu Gln Val Lys Asp Ile Thr Glu Asn Me - #t Leu Cys Ala Gly Val               35         - #         40         - #         45                      - - Arg Arg Gly Gly Lys Asp Ser Cys                                               50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:22:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 177 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..177                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                              - - GGA TGG GGA TCA AGA TCT ACT TCC AAT TTC CC - #A TCT TAC CCC AAC CTT           48                                                                       Gly Trp Gly Ser Arg Ser Thr Ser Asn Phe Pr - #o Ser Tyr Pro Asn Leu             1               5 - #                 10 - #                 15              - - TTA CAG ACC GTT GAC AAA CCA ATT GTA TCT TA - #T GCC GAA TGT GAG AAA           96                                                                       Leu Gln Thr Val Asp Lys Pro Ile Val Ser Ty - #r Ala Glu Cys Glu Lys                        20     - #             25     - #             30                  - - GTA TTG GGA GGT CCT GGA GCC TCA CCA CTT CA - #C CCC TTG AAC CTC TGC          144                                                                       Val Leu Gly Gly Pro Gly Ala Ser Pro Leu Hi - #s Pro Leu Asn Leu Cys                    35         - #         40         - #         45                      - - ACT GGA CCC TTG ACC GGT GGA GTA AGC GCT TG - #T                  -      #        177                                                                    Thr Gly Pro Leu Thr Gly Gly Val Ser Ala Cy - #s                                    50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:23:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 59 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                              - - Gly Trp Gly Ser Arg Ser Thr Ser Asn Phe Pr - #o Ser Tyr Pro Asn        Leu                                                                               1               5 - #                 10 - #                 15             - - Leu Gln Thr Val Asp Lys Pro Ile Val Ser Ty - #r Ala Glu Cys Glu Lys                   20     - #             25     - #             30                  - - Val Leu Gly Gly Pro Gly Ala Ser Pro Leu Hi - #s Pro Leu Asn Leu Cys               35         - #         40         - #         45                      - - Thr Gly Pro Leu Thr Gly Gly Val Ser Ala Cy - #s                               50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:24:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 156 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..156                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                              - - GGC TGG GGA AAT ACA AAT TCA GCA GGG GAA AA - #T TTT CCA TTG AAA CTT           48                                                                       Gly Trp Gly Asn Thr Asn Ser Ala Gly Glu As - #n Phe Pro Leu Lys Leu             1               5 - #                 10 - #                 15              - - CAA GAA TTG TAC GTG AAA GCT TTG ACT AAT GA - #G GAG TGC AAA GCT AAA           96                                                                       Gln Glu Leu Tyr Val Lys Ala Leu Thr Asn Gl - #u Glu Cys Lys Ala Lys                        20     - #             25     - #             30                  - - TCA CCA ATT CCA CCA ACG ACC CAA GTC TGC AC - #A CTT TTG GAA AAG AAT          144                                                                       Ser Pro Ile Pro Pro Thr Thr Gln Val Cys Th - #r Leu Leu Glu Lys Asn                    35         - #         40         - #         45                      - - CAT GGT GTA TGC            - #                  - #                      - #      156                                                                  His Gly Val Cys                                                                    50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:25:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 52 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                              - - Gly Trp Gly Asn Thr Asn Ser Ala Gly Glu As - #n Phe Pro Leu Lys Leu        1               5 - #                 10 - #                 15              - - Gln Glu Leu Tyr Val Lys Ala Leu Thr Asn Gl - #u Glu Cys Lys Ala Lys                   20     - #             25     - #             30                  - - Ser Pro Ile Pro Pro Thr Thr Gln Val Cys Th - #r Leu Leu Glu Lys Asn               35         - #         40         - #         45                      - - His Gly Val Cys                                                               50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:26:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 159 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..159                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                              - - GGA TGG GGA TCA ACT GGA TCT GGT GGT CCA AT - #T ACA AAT GTT CTA CAA           48                                                                       Gly Trp Gly Ser Thr Gly Ser Gly Gly Pro Il - #e Thr Asn Val Leu Gln             1               5 - #                 10 - #                 15              - - GAA GTC GAA GTT CCA TTT ATC GAC TTC AAC AC - #C TGC CGA AAA TCC TAC           96                                                                       Glu Val Glu Val Pro Phe Ile Asp Phe Asn Th - #r Cys Arg Lys Ser Tyr                        20     - #             25     - #             30                  - - TCA ACC AGC TTA ACC GAC CGT ATG TTC TGC GC - #T GGA TTT TTG GGA ATT          144                                                                       Ser Thr Ser Leu Thr Asp Arg Met Phe Cys Al - #a Gly Phe Leu Gly Ile                    35         - #         40         - #         45                      - - GGT GGT AAG GCT TGC           - #                  - #                      - #   159                                                                  Gly Gly Lys Ala Cys                                                                50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:27:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 53 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                              - - Gly Trp Gly Ser Thr Gly Ser Gly Gly Pro Il - #e Thr Asn Val Leu Gln        1               5 - #                 10 - #                 15              - - Glu Val Glu Val Pro Phe Ile Asp Phe Asn Th - #r Cys Arg Lys Ser Tyr                   20     - #             25     - #             30                  - - Ser Thr Ser Leu Thr Asp Arg Met Phe Cys Al - #a Gly Phe Leu Gly Ile               35         - #         40         - #         45                      - - Gly Gly Lys Ala Cys                                                           50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:28:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 168 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..168                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                              - - GGC TGG GGA AAT TTA GGG GAA GAT GAG GAC GA - #C CCC GAA CAA CTG CAA           48                                                                       Gly Trp Gly Asn Leu Gly Glu Asp Glu Asp As - #p Pro Glu Gln Leu Gln             1               5 - #                 10 - #                 15              - - TAT GTA AAG GTA CCT ATT GTT AAC TGG ACT CA - #G TGC AAA ACT ATA TAT           96                                                                       Tyr Val Lys Val Pro Ile Val Asn Trp Thr Gl - #n Cys Lys Thr Ile Tyr                        20     - #             25     - #             30                  - - GGA AAT GAA GGA CTA ATA ATT ACC CAA AAT AT - #G ATT TGT GCT GGT TAT          144                                                                       Gly Asn Glu Gly Leu Ile Ile Thr Gln Asn Me - #t Ile Cys Ala Gly Tyr                    35         - #         40         - #         45                      - - CCT GAT GGC GGT AAG GAC TCT TGC     - #                  - #                   168                                                                     Pro Asp Gly Gly Lys Asp Ser Cys                                                    50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:29:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 56 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                              - - Gly Trp Gly Asn Leu Gly Glu Asp Glu Asp As - #p Pro Glu Gln Leu Gln        1               5 - #                 10 - #                 15              - - Tyr Val Lys Val Pro Ile Val Asn Trp Thr Gl - #n Cys Lys Thr Ile Tyr                   20     - #             25     - #             30                  - - Gly Asn Glu Gly Leu Ile Ile Thr Gln Asn Me - #t Ile Cys Ala Gly Tyr               35         - #         40         - #         45                      - - Pro Asp Gly Gly Lys Asp Ser Cys                                               50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:30:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 159 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..159                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                              - - GGA TGG GCA TCT CCA AAG ATT TCC CCT GCT TT - #C GAA TTG CCT GAC AAA           48                                                                       Gly Trp Ala Ser Pro Lys Ile Ser Pro Ala Ph - #e Glu Leu Pro Asp Lys             1               5 - #                 10 - #                 15              - - CTA CAG TAC ACA ACT TTG GAA GTC CAA CCA AG - #T GAA GAC TGC AAA AAA           96                                                                       Leu Gln Tyr Thr Thr Leu Glu Val Gln Pro Se - #r Glu Asp Cys Lys Lys                        20     - #             25     - #             30                  - - GTA TGG GCC CCT TAC ATG CGC GAC TAC ATC CT - #T TGT GCC AAA TTT GAA          144                                                                       Val Trp Ala Pro Tyr Met Arg Asp Tyr Ile Le - #u Cys Ala Lys Phe Glu                    35         - #         40         - #         45                      - - AAA CAA AAC ATT TGC           - #                  - #                      - #   159                                                                  Lys Gln Asn Ile Cys                                                                50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:31:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 53 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                              - - Gly Trp Ala Ser Pro Lys Ile Ser Pro Ala Ph - #e Glu Leu Pro Asp Lys        1               5 - #                 10 - #                 15              - - Leu Gln Tyr Thr Thr Leu Glu Val Gln Pro Se - #r Glu Asp Cys Lys Lys                   20     - #             25     - #             30                  - - Val Trp Ala Pro Tyr Met Arg Asp Tyr Ile Le - #u Cys Ala Lys Phe Glu               35         - #         40         - #         45                      - - Lys Gln Asn Ile Cys                                                           50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:32:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 186 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..186                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 86/87, change                         A/A to - #V/Y; at pos. aa 29, change Glu to Xaa and -         #define                                                                                        Xaa = - #Val, Ala, Asp, Glu or Gly. "                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                              - - GGT TGG GGA AAG ATA GAC TAT TCT GAG AGC AG - #A AGT GAT GAC CTA        CTG       48                                                                    Gly Trp Gly Lys Ile Asp Tyr Ser Glu Ser Ar - #g Ser Asp Asp Leu Leu            1               5 - #                 10 - #                 15              - - AAA GTA GTA CTG AAA ATT ATT GAT AAT AGG CA - #A TGC GVY CCC TTA TAC           96                                                                       Lys Val Val Leu Lys Ile Ile Asp Asn Arg Gl - #n Cys Xaa Pro Leu Tyr                        20     - #             25     - #             30                  - - GTT GAT CAG ATT AAT AGA AGA AGA TTG AGA AA - #T GGA ATT GTA GAA ACA          144                                                                       Val Asp Gln Ile Asn Arg Arg Arg Leu Arg As - #n Gly Ile Val Glu Thr                    35         - #         40         - #         45                      - - CAG ATG TGT GCA GGA GAA TTG GAT GGT GGA AA - #A GAC ACT TGC                 - # 186                                                                    Gln Met Cys Ala Gly Glu Leu Asp Gly Gly Ly - #s Asp Thr Cys                        50             - #     55             - #     60                          - -  - - (2) INFORMATION FOR SEQ ID NO:33:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 62 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (ix) FEATURE:                                                                  (A) NAME/KEY: Xaa = - #Val, Ala, Asp Glu or Gly                               (B) LOCATION: 29                                                     - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                              - - Gly Trp Gly Lys Ile Asp Tyr Ser Glu Ser Ar - #g Ser Asp Asp Leu Leu        1               5 - #                 10 - #                 15              - - Lys Val Val Leu Lys Ile Ile Asp Asn Arg Gl - #n Cys Xaa Pro Leu Tyr                   20     - #             25     - #             30                  - - Val Asp Gln Ile Asn Arg Arg Arg Leu Arg As - #n Gly Ile Val Glu Thr               35         - #         40         - #         45                      - - Gln Met Cys Ala Gly Glu Leu Asp Gly Gly Ly - #s Asp Thr Cys                   50             - #     55             - #     60                          - -  - - (2) INFORMATION FOR SEQ ID NO:34:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 168 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..168                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                              - - GGA TGG GGA AGA ACA TCG TTC GGT GGC CAA TT - #G TCT AAA AAT CTG CGA           48                                                                       Gly Trp Gly Arg Thr Ser Phe Gly Gly Gln Le - #u Ser Lys Asn Leu Arg             1               5 - #                 10 - #                 15              - - GGA GTC GAG TTG GAA ATA ATA GAT CTA TTC GA - #T TGT TTC CTT TCC TAC           96                                                                       Gly Val Glu Leu Glu Ile Ile Asp Leu Phe As - #p Cys Phe Leu Ser Tyr                        20     - #             25     - #             30                  - - ATG GAT AAA GTA AAC GTG TCC GAA AGG CAA GT - #T TGC GCT GGA ATC CCC          144                                                                       Met Asp Lys Val Asn Val Ser Glu Arg Gln Va - #l Cys Ala Gly Ile Pro                    35         - #         40         - #         45                      - - GTT GTA GGT GGT AAA GAT TCT TGC     - #                  - #                   168                                                                     Val Val Gly Gly Lys Asp Ser Cys                                                    50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:35:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 56 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                              - - Gly Trp Gly Arg Thr Ser Phe Gly Gly Gln Le - #u Ser Lys Asn Leu Arg        1               5 - #                 10 - #                 15              - - Gly Val Glu Leu Glu Ile Ile Asp Leu Phe As - #p Cys Phe Leu Ser Tyr                   20     - #             25     - #             30                  - - Met Asp Lys Val Asn Val Ser Glu Arg Gln Va - #l Cys Ala Gly Ile Pro               35         - #         40         - #         45                      - - Val Val Gly Gly Lys Asp Ser Cys                                               50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:36:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 120 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..120                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                              - - GGA TGG GGT GCA GTC TAC GAA GGA GGT GCA GG - #A TCC ACC CAA TTA CTA           48                                                                       Gly Trp Gly Ala Val Tyr Glu Gly Gly Ala Gl - #y Ser Thr Gln Leu Leu             1               5 - #                 10 - #                 15              - - TAC TCC CAA TTT GGC GGT GTT GCT CCT AGC AT - #G ATC TGC GCT GGA TTT           96                                                                       Tyr Ser Gln Phe Gly Gly Val Ala Pro Ser Me - #t Ile Cys Ala Gly Phe                        20     - #             25     - #             30                  - - GAC CAA GGC GGT AAG GAC GCT TGT     - #                  - #                   120                                                                     Asp Gln Gly Gly Lys Asp Ala Cys                                                        35         - #         40                                             - -  - - (2) INFORMATION FOR SEQ ID NO:37:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 40 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                              - - Gly Trp Gly Ala Val Tyr Glu Gly Gly Ala Gl - #y Ser Thr Gln Leu Leu        1               5 - #                 10 - #                 15              - - Tyr Ser Gln Phe Gly Gly Val Ala Pro Ser Me - #t Ile Cys Ala Gly Phe                   20     - #             25     - #             30                  - - Asp Gln Gly Gly Lys Asp Ala Cys                                                   35         - #         40                                             - -  - - (2) INFORMATION FOR SEQ ID NO:38:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 162 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..162                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                              - - GGT TGG GGA ACT ACA GAG AGT ACT GAA TCA TC - #A CAC CAC CTG AAA GAA           48                                                                       Gly Trp Gly Thr Thr Glu Ser Thr Glu Ser Se - #r His His Leu Lys Glu             1               5 - #                 10 - #                 15              - - GTT GAA GTG AAC GCT GTA TCT AAT AGT GAA TG - #T CAA AGG CCT AAT GAA           96                                                                       Val Glu Val Asn Ala Val Ser Asn Ser Glu Cy - #s Gln Arg Pro Asn Glu                        20     - #             25     - #             30                  - - GAT CTT GCT ACT ATA TCA TCA CAT GAG ATA TG - #T GCA AGC GTT CCT GGT          144                                                                       Asp Leu Ala Thr Ile Ser Ser His Glu Ile Cy - #s Ala Ser Val Pro Gly                    35         - #         40         - #         45                      - - GGC GGC AAA GAT TCT TGT         - #                  - #                      - # 162                                                                  Gly Gly Lys Asp Ser Cys                                                            50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:39:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 54 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                              - - Gly Trp Gly Thr Thr Glu Ser Thr Glu Ser Se - #r His His Leu Lys Glu        1               5 - #                 10 - #                 15              - - Val Glu Val Asn Ala Val Ser Asn Ser Glu Cy - #s Gln Arg Pro Asn Glu                   20     - #             25     - #             30                  - - Asp Leu Ala Thr Ile Ser Ser His Glu Ile Cy - #s Ala Ser Val Pro Gly               35         - #         40         - #         45                      - - Gly Gly Lys Asp Ser Cys                                                       50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:40:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 40 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                              - - Ile Val Gly Gly Glu Asn Ala Lys Glu Lys Se - #r Asp Val Pro Tyr Gln      1               5   - #                10  - #                15               - - Val Ser Leu Arg Asn Ala Glu Asn Lys His Ph - #e Cys Gly Gly Ala Ile                  20      - #            25      - #            30                   - - Ile Asp Asp Tyr Trp Val Leu Thr                                                  35          - #        40                                              - -  - - (2) INFORMATION FOR SEQ ID NO:41:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 25 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                              - - Ile Val Gly Gly Leu Glu Ala Lys Asn Gly Se - #r Ala Pro Phe Met Val      1               5   - #                10  - #                15               - - Ser Leu Gln Ala Glu Asp Tyr Phe His                                                  20      - #            25                                          - -  - - (2) INFORMATION FOR SEQ ID NO:42:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 26 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                              - - Ile Ile Gly Gly Glu Val Ala Gly Glu Gly Se - #r Ala Pro Tyr Gln Val      1               5   - #                10  - #                15               - - Ser Leu Arg Thr Lys Glu Gly Asn His Phe                                              20      - #            25                                          - -  - - (2) INFORMATION FOR SEQ ID NO:43:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                              - - Ile Val Gly Gly Thr Ala Val Asp Ile Arg Gl - #y Phe Pro Gly Arg Tyr      1               5   - #                10  - #                15               - - Gln Phe Lys Pro Lys Pro Ser Phe Leu Trp Tr - #p Phe Tyr                              20      - #            25                                          - -  - - (2) INFORMATION FOR SEQ ID NO:44:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 35 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                              - - Ile Val Asn Gly Leu Glu Ala Gly Val Gly Gl - #n Phe Pro Ile Gln Val      1               5   - #                10  - #                15               - - Phe Leu Asp Leu Thr Asn Ile Arg Asp Glu Ly - #s Ser Arg Cys Gly Gly                  20      - #            25      - #            30                   - - Ala Leu Phe                                                                      35                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:45:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                              - - Ile Val Gly Gly Leu Glu Ala Lys Asn Gly Il - #e Thr Pro Phe Ile Gly      1               5   - #                10  - #                15               - - Phe Phe Ala Ser Gly Arg Leu Phe                                                      20                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:46:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 26 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                              - - Ile Val Gly Gly Asn Asp Val Ser Xaa Lys Il - #e Phe Trp Gln Val Ser      1               5   - #                10  - #                15               - - Ile Gln Ser Asn Xaa Gln His Phe Cys Gly                                              20      - #            25                                          - -  - - (2) INFORMATION FOR SEQ ID NO:47:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                              - - Ile Ile Gly Gly Glu Asp Ala Pro Glu Gly Se - #r Ala Pro Tyr Gln Val      1               5   - #                10  - #                15               - - Ser Leu Arg Asn Gln Asn Leu Glu His Phe Cy - #s Gly Gly Ser Ile                      20      - #            25      - #            30                   - -  - - (2) INFORMATION FOR SEQ ID NO:48:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 32 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                              - - GTWGGWAAAG GWWTWACWTT YGATTCWGGW GG       - #                  - #              32                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:49:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                              - - CGWCCTTCWG CATCWGTATT            - #                  - #                      - # 20                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:50:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 453 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..453                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                              - - CAC GAG TTT TGT GCG AGT GTC AGA TAT TGC AG - #C TCT ATG AGT AAC AAG           48                                                                       His Glu Phe Cys Ala Ser Val Arg Tyr Cys Se - #r Ser Met Ser Asn Lys             1               5 - #                 10 - #                 15              - - AAA GGA TTA GTA CTG GGC ATC TAC GAC AAT GA - #A TTC GAT AAA AAA ATA           96                                                                       Lys Gly Leu Val Leu Gly Ile Tyr Asp Asn Gl - #u Phe Asp Lys Lys Ile                        20     - #             25     - #             30                  - - AGG TTA ACG CCA ACT GCT GAA CAA TTC AAT CG - #G CGA TTG CAG GGG CGT          144                                                                       Arg Leu Thr Pro Thr Ala Glu Gln Phe Asn Ar - #g Arg Leu Gln Gly Arg                    35         - #         40         - #         45                      - - TTA CTA GAT CTA ATT CAT TTG AGT GGA CCC AT - #T AAA TTG GGC AAG AGC          192                                                                       Leu Leu Asp Leu Ile His Leu Ser Gly Pro Il - #e Lys Leu Gly Lys Ser                50             - #     55             - #     60                          - - CGT ATT TTC TGG GAT CTC GAT GAA TTC GGC GC - #A GTT GCA GTT GCA GGT          240                                                                       Arg Ile Phe Trp Asp Leu Asp Glu Phe Gly Al - #a Val Ala Val Ala Gly            65                 - # 70                 - # 75                 - # 80       - - TTG GGA AAT CAC TCC CCC TGC GAA CTC CTG GA - #A GAA CTC GAT GTT TTG          288                                                                       Leu Gly Asn His Ser Pro Cys Glu Leu Leu Gl - #u Glu Leu Asp Val Leu                            85 - #                 90 - #                 95              - - CGC GAA AAT GCC AGA ATA GCT GCC GGT GCT GG - #T TGC CAA GCT CTT GCC          336                                                                       Arg Glu Asn Ala Arg Ile Ala Ala Gly Ala Gl - #y Cys Gln Ala Leu Ala                       100      - #           105      - #           110                  - - GCC GAT GGA ATC ACT ACC ATT AGC GTT GAA GT - #A TGG AGC ACC CGG AGG          384                                                                       Ala Asp Gly Ile Thr Thr Ile Ser Val Glu Va - #l Trp Ser Thr Arg Arg                   115          - #       120          - #       125                      - - CGG CCA TGC GAA GGT GCA ATA CTA TCG ACG TT - #C AAA TTC AGG TCA ACA          432                                                                       Arg Pro Cys Glu Gly Ala Ile Leu Ser Thr Ph - #e Lys Phe Arg Ser Thr               130              - #   135              - #   140                          - - GAA GTA GTC CAG TGT AGC GGT       - #                  - #                     453                                                                     Glu Val Val Gln Cys Ser Gly                                                   145                 1 - #50                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:51:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 151 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                              - - His Glu Phe Cys Ala Ser Val Arg Tyr Cys Se - #r Ser Met Ser Asn Lys        1               5 - #                 10 - #                 15              - - Lys Gly Leu Val Leu Gly Ile Tyr Asp Asn Gl - #u Phe Asp Lys Lys Ile                   20     - #             25     - #             30                  - - Arg Leu Thr Pro Thr Ala Glu Gln Phe Asn Ar - #g Arg Leu Gln Gly Arg               35         - #         40         - #         45                      - - Leu Leu Asp Leu Ile His Leu Ser Gly Pro Il - #e Lys Leu Gly Lys Ser           50             - #     55             - #     60                          - - Arg Ile Phe Trp Asp Leu Asp Glu Phe Gly Al - #a Val Ala Val Ala Gly       65                 - # 70                 - # 75                 - # 80       - - Leu Gly Asn His Ser Pro Cys Glu Leu Leu Gl - #u Glu Leu Asp Val Leu                       85 - #                 90 - #                 95              - - Arg Glu Asn Ala Arg Ile Ala Ala Gly Ala Gl - #y Cys Gln Ala Leu Ala                  100      - #           105      - #           110                  - - Ala Asp Gly Ile Thr Thr Ile Ser Val Glu Va - #l Trp Ser Thr Arg Arg              115          - #       120          - #       125                      - - Arg Pro Cys Glu Gly Ala Ile Leu Ser Thr Ph - #e Lys Phe Arg Ser Thr          130              - #   135              - #   140                          - - Glu Val Val Gln Cys Ser Gly                                              145                 1 - #50                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:52:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 258 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..258                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                              - - TCA GCA CTC GTT GCC TTG TCT GCA GCT ATT CC - #T CAC TCC AAC AGA GTC           48                                                                       Ser Ala Leu Val Ala Leu Ser Ala Ala Ile Pr - #o His Ser Asn Arg Val             1               5 - #                 10 - #                 15              - - GTT GGA GGA CTG GAA GCT GCA GAG GGT TCT GC - #A CCT TAT CAA GTA TCC           96                                                                       Val Gly Gly Leu Glu Ala Ala Glu Gly Ser Al - #a Pro Tyr Gln Val Ser                        20     - #             25     - #             30                  - - TTG CAA GTT GGC AAC TTC CAC TTC TGT GGT GG - #T TCA ATT CTG AAC GAA          144                                                                       Leu Gln Val Gly Asn Phe His Phe Cys Gly Gl - #y Ser Ile Leu Asn Glu                    35         - #         40         - #         45                      - - TAT TGG GTT TTG ACT GCT GCT CAC TGT TTG GG - #T TAT GAC TTC GAC GTG          192                                                                       Tyr Trp Val Leu Thr Ala Ala His Cys Leu Gl - #y Tyr Asp Phe Asp Val                50             - #     55             - #     60                          - - GTA GTT GGA ACA AAC AAA CTT GAT CAA CCA GG - #T GAA AGA TAC CTC GTA          240                                                                       Val Val Gly Thr Asn Lys Leu Asp Gln Pro Gl - #y Glu Arg Tyr Leu Val            65                 - # 70                 - # 75                 - # 80       - - GAA CAA ACT TTT GTT CAC         - #                  - #                      - # 258                                                                  Glu Gln Thr Phe Val His                                                                        85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:53:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 86 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                              - - Ser Ala Leu Val Ala Leu Ser Ala Ala Ile Pr - #o His Ser Asn Arg Val        1               5 - #                 10 - #                 15              - - Val Gly Gly Leu Glu Ala Ala Glu Gly Ser Al - #a Pro Tyr Gln Val Ser                   20     - #             25     - #             30                  - - Leu Gln Val Gly Asn Phe His Phe Cys Gly Gl - #y Ser Ile Leu Asn Glu               35         - #         40         - #         45                      - - Tyr Trp Val Leu Thr Ala Ala His Cys Leu Gl - #y Tyr Asp Phe Asp Val           50             - #     55             - #     60                          - - Val Val Gly Thr Asn Lys Leu Asp Gln Pro Gl - #y Glu Arg Tyr Leu Val       65                 - # 70                 - # 75                 - # 80       - - Glu Gln Thr Phe Val His                                                                   85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:54:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 240 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..240                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                              - - TTA GAT GGG CGC ATT GTT GGA GGA CAA GAT GC - #T GAT ATT GCC AAA TAT           48                                                                       Leu Asp Gly Arg Ile Val Gly Gly Gln Asp Al - #a Asp Ile Ala Lys Tyr             1               5 - #                 10 - #                 15              - - GGC TAT CAA GCT TCA CTC CAA GTA TTT AAC GA - #A CAT TTC TGT GGA GCT           96                                                                       Gly Tyr Gln Ala Ser Leu Gln Val Phe Asn Gl - #u His Phe Cys Gly Ala                        20     - #             25     - #             30                  - - TCA ATA TTG AAT AAT TAT TGG ATT GTC ACA GC - #A GCT CAT TGC ATA TAT          144                                                                       Ser Ile Leu Asn Asn Tyr Trp Ile Val Thr Al - #a Ala His Cys Ile Tyr                    35         - #         40         - #         45                      - - GAT GAA TTC ACG TAT TCA GTT CGA GTC GGC AC - #C AGT TTC CAA GGA AGA          192                                                                       Asp Glu Phe Thr Tyr Ser Val Arg Val Gly Th - #r Ser Phe Gln Gly Arg                50             - #     55             - #     60                          - - CGT GGT TCC GTT CAT CCT GTG GCA CAA ATT AT - #C AAG CAT CCT GCA TAC          240                                                                       Arg Gly Ser Val His Pro Val Ala Gln Ile Il - #e Lys His Pro Ala Tyr            65                 - # 70                 - # 75                 - # 80       - -  - - (2) INFORMATION FOR SEQ ID NO:55:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 80 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                              - - Leu Asp Gly Arg Ile Val Gly Gly Gln Asp Al - #a Asp Ile Ala Lys Tyr        1               5 - #                 10 - #                 15              - - Gly Tyr Gln Ala Ser Leu Gln Val Phe Asn Gl - #u His Phe Cys Gly Ala                   20     - #             25     - #             30                  - - Ser Ile Leu Asn Asn Tyr Trp Ile Val Thr Al - #a Ala His Cys Ile Tyr               35         - #         40         - #         45                      - - Asp Glu Phe Thr Tyr Ser Val Arg Val Gly Th - #r Ser Phe Gln Gly Arg           50             - #     55             - #     60                          - - Arg Gly Ser Val His Pro Val Ala Gln Ile Il - #e Lys His Pro Ala Tyr       65                 - # 70                 - # 75                 - # 80       - -  - - (2) INFORMATION FOR SEQ ID NO:56:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 218 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..216                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                              - - AGG GAA CAA AAG CTG GAG CTC CAC CGC GGT GC - #G CCG GCT CTA GAA CTA           48                                                                       Arg Glu Gln Lys Leu Glu Leu His Arg Gly Al - #a Pro Ala Leu Glu Leu             1               5 - #                 10 - #                 15              - - GTG GAT CCC CCG GGT CTG CAG GAA TTG GCA CG - #A GGA TGT TCT TGG CTG           96                                                                       Val Asp Pro Pro Gly Leu Gln Glu Leu Ala Ar - #g Gly Cys Ser Trp Leu                        20     - #             25     - #             30                  - - TGT TTA GTA GCT ATT CTT TGT GCA GTG GCT GC - #T GGG CCT ACT AAT CGC          144                                                                       Cys Leu Val Ala Ile Leu Cys Ala Val Ala Al - #a Gly Pro Thr Asn Arg                    35         - #         40         - #         45                      - - ATT GTT GGA GGA TTG GAG GCG AAA AAT GGA AT - #C ACC CCA TTC ATC GGT          192                                                                       Ile Val Gly Gly Leu Glu Ala Lys Asn Gly Il - #e Thr Pro Phe Ile Gly                50             - #     55             - #     60                          - - TTC TTT GCA AGC GGA AGA CTA TTT CA    - #                  - #                 218                                                                     Phe Phe Ala Ser Gly Arg Leu Phe                                                65                 - # 70                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:57:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 72 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                              - - Arg Glu Gln Lys Leu Glu Leu His Arg Gly Al - #a Pro Ala Leu Glu Leu        1               5 - #                 10 - #                 15              - - Val Asp Pro Pro Gly Leu Gln Glu Leu Ala Ar - #g Gly Cys Ser Trp Leu                   20     - #             25     - #             30                  - - Cys Leu Val Ala Ile Leu Cys Ala Val Ala Al - #a Gly Pro Thr Asn Arg               35         - #         40         - #         45                      - - Ile Val Gly Gly Leu Glu Ala Lys Asn Gly Il - #e Thr Pro Phe Ile Gly           50             - #     55             - #     60                          - - Phe Phe Ala Ser Gly Arg Leu Phe                                           65                 - # 70                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:58:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 240 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..240                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                              - - ACG AGG TTT CGC TTA GCA ATT GTA TGT GCT CT - #C GCT GTC TGC ACA TTC           48                                                                       Thr Arg Phe Arg Leu Ala Ile Val Cys Ala Le - #u Ala Val Cys Thr Phe             1               5 - #                 10 - #                 15              - - GGT GCC AGT GTT CCA GAA CCA TGG AAA AGA TT - #A GAT GGT AGA ATC GTA           96                                                                       Gly Ala Ser Val Pro Glu Pro Trp Lys Arg Le - #u Asp Gly Arg Ile Val                        20     - #             25     - #             30                  - - GGA GGA CAC GAT ACC AGC ATC GAT AAA CAC CC - #T CAT CAA GTA TCT TTA          144                                                                       Gly Gly His Asp Thr Ser Ile Asp Lys His Pr - #o His Gln Val Ser Leu                    35         - #         40         - #         45                      - - TTG TAC TCC AGC CAC AAT TGT GGT GGT TCC TT - #G ATT GCC AAA AAC TGG          192                                                                       Leu Tyr Ser Ser His Asn Cys Gly Gly Ser Le - #u Ile Ala Lys Asn Trp                50             - #     55             - #     60                          - - GTT TTG ACT GCA GCT CAT TGC ATT GGA GTT AA - #C AAA TAC AAT GTC CGT          240                                                                       Val Leu Thr Ala Ala His Cys Ile Gly Val As - #n Lys Tyr Asn Val Arg            65                 - # 70                 - # 75                 - # 80       - -  - - (2) INFORMATION FOR SEQ ID NO:59:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 80 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                              - - Thr Arg Phe Arg Leu Ala Ile Val Cys Ala Le - #u Ala Val Cys Thr Phe        1               5 - #                 10 - #                 15              - - Gly Ala Ser Val Pro Glu Pro Trp Lys Arg Le - #u Asp Gly Arg Ile Val                   20     - #             25     - #             30                  - - Gly Gly His Asp Thr Ser Ile Asp Lys His Pr - #o His Gln Val Ser Leu               35         - #         40         - #         45                      - - Leu Tyr Ser Ser His Asn Cys Gly Gly Ser Le - #u Ile Ala Lys Asn Trp           50             - #     55             - #     60                          - - Val Leu Thr Ala Ala His Cys Ile Gly Val As - #n Lys Tyr Asn Val Arg       65                 - # 70                 - # 75                 - # 80       - -  - - (2) INFORMATION FOR SEQ ID NO:60:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 234 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..234                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                              - - CCC TCA CTA AAG GGA ACA AAA GCT GGA GCT CC - #A CCG CGG TGC GCC GCT           48                                                                       Pro Ser Leu Lys Gly Thr Lys Ala Gly Ala Pr - #o Pro Arg Cys Ala Ala             1               5 - #                 10 - #                 15              - - CTA GAA CTA GTG GAT CCC CCG GGC TGC AGG AA - #T TCG GCA CGA GCG TTT           96                                                                       Leu Glu Leu Val Asp Pro Pro Gly Cys Arg As - #n Ser Ala Arg Ala Phe                        20     - #             25     - #             30                  - - GGT TGG ATT GAG CGC GTC TCA TCT TAC AAG AT - #A AAG GAT AGA TTA GAT          144                                                                       Gly Trp Ile Glu Arg Val Ser Ser Tyr Lys Il - #e Lys Asp Arg Leu Asp                    35         - #         40         - #         45                      - - GGG CGC ATT GTT GGA GGA CAA GAT GCT GAT AT - #T GCC AAA TAT GGC TAT          192                                                                       Gly Arg Ile Val Gly Gly Gln Asp Ala Asp Il - #e Ala Lys Tyr Gly Tyr                50             - #     55             - #     60                          - - CAA GCT TCA CTC CAA GTA CTT AAC GAA CAT TT - #C TGT GGA GCT                 - # 234                                                                    Gln Ala Ser Leu Gln Val Leu Asn Glu His Ph - #e Cys Gly Ala                    65                 - # 70                 - # 75                              - -  - - (2) INFORMATION FOR SEQ ID NO:61:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 78 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                              - - Pro Ser Leu Lys Gly Thr Lys Ala Gly Ala Pr - #o Pro Arg Cys Ala Ala        1               5 - #                 10 - #                 15              - - Leu Glu Leu Val Asp Pro Pro Gly Cys Arg As - #n Ser Ala Arg Ala Phe                   20     - #             25     - #             30                  - - Gly Trp Ile Glu Arg Val Ser Ser Tyr Lys Il - #e Lys Asp Arg Leu Asp               35         - #         40         - #         45                      - - Gly Arg Ile Val Gly Gly Gln Asp Ala Asp Il - #e Ala Lys Tyr Gly Tyr           50             - #     55             - #     60                          - - Gln Ala Ser Leu Gln Val Leu Asn Glu His Ph - #e Cys Gly Ala               65                 - # 70                 - # 75                              - -  - - (2) INFORMATION FOR SEQ ID NO:62:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 291 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..291                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                              - - GCG GTG ATT GTG TCA TTT GTT CTG GCT TGT GC - #A TTT TCT GTA CAG GCT           48                                                                       Ala Val Ile Val Ser Phe Val Leu Ala Cys Al - #a Phe Ser Val Gln Ala             1               5 - #                 10 - #                 15              - - CTT CCA TCA AGC AGA ATT GTC AAT GGA CTT GA - #A GCA GGA GTT GGA CAA           96                                                                       Leu Pro Ser Ser Arg Ile Val Asn Gly Leu Gl - #u Ala Gly Val Gly Gln                        20     - #             25     - #             30                  - - TTT CCA ATT CAG GTT TTC TTA GAC TTG ACA AA - #T ATC AGA GAC GAA AAA          144                                                                       Phe Pro Ile Gln Val Phe Leu Asp Leu Thr As - #n Ile Arg Asp Glu Lys                    35         - #         40         - #         45                      - - TCC AGA TGT GGT GGT GCT TTG TTA TCA GAT TC - #A TGG GTT TTG ACT GCT          192                                                                       Ser Arg Cys Gly Gly Ala Leu Leu Ser Asp Se - #r Trp Val Leu Thr Ala                50             - #     55             - #     60                          - - GCT CAT TGT TTT GAT GAT TTG AAG TCT ATG GT - #A GTG TCC GTT GGT GCT          240                                                                       Ala His Cys Phe Asp Asp Leu Lys Ser Met Va - #l Val Ser Val Gly Ala            65                 - # 70                 - # 75                 - # 80       - - CAT GAT GTC AGC AAA TCT GAA GAA CCT CAC AG - #G CAA ACC AGG AAA CCT          288                                                                       His Asp Val Ser Lys Ser Glu Glu Pro His Ar - #g Gln Thr Arg Lys Pro                            85 - #                 90 - #                 95              - - GAA                  - #                  - #                  - #                291                                                                  Glu                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:63:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 97 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                              - - Ala Val Ile Val Ser Phe Val Leu Ala Cys Al - #a Phe Ser Val Gln Ala        1               5 - #                 10 - #                 15              - - Leu Pro Ser Ser Arg Ile Val Asn Gly Leu Gl - #u Ala Gly Val Gly Gln                   20     - #             25     - #             30                  - - Phe Pro Ile Gln Val Phe Leu Asp Leu Thr As - #n Ile Arg Asp Glu Lys               35         - #         40         - #         45                      - - Ser Arg Cys Gly Gly Ala Leu Leu Ser Asp Se - #r Trp Val Leu Thr Ala           50             - #     55             - #     60                          - - Ala His Cys Phe Asp Asp Leu Lys Ser Met Va - #l Val Ser Val Gly Ala       65                 - # 70                 - # 75                 - # 80       - - His Asp Val Ser Lys Ser Glu Glu Pro His Ar - #g Gln Thr Arg Lys Pro                       85 - #                 90 - #                 95              - - Glu                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:64:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 144 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..144                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 108, change A to                      M; at - #pos. 135, change C to Y; at pos. 141, change - #     A to                                                                                          R; at - #pos. 144, change A to R."                              - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                              - - GTA CTG ATC GTT TTA GCA GTC ATT GAA TTC GC - #A TCA GCG TCT TCA ATC           48                                                                       Val Leu Ile Val Leu Ala Val Ile Glu Phe Al - #a Ser Ala Ser Ser Ile             1               5 - #                 10 - #                 15              - - GGC TGG AGA ATC GTG GGT GGT GAA AAT GCT AA - #A GAA AAA TCG GTG CCC           96                                                                       Gly Trp Arg Ile Val Gly Gly Glu Asn Ala Ly - #s Glu Lys Ser Val Pro                        20     - #             25     - #             30                  - - TAT CAA GTT TCM CTT CGA AAT GCT GAA AAC AA - #A CAT TTY TGT GGR GGR          144                                                                       Tyr Gln Val Ser Leu Arg Asn Ala Glu Asn Ly - #s His Phe Cys Gly Gly                    35         - #         40         - #         45                      - -  - - (2) INFORMATION FOR SEQ ID NO:65:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 48 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                              - - Val Leu Ile Val Leu Ala Val Ile Glu Phe Al - #a Ser Ala Ser Ser Ile        1               5 - #                 10 - #                 15              - - Gly Trp Arg Ile Val Gly Gly Glu Asn Ala Ly - #s Glu Lys Ser Val Pro                   20     - #             25     - #             30                  - - Tyr Gln Val Ser Leu Arg Asn Ala Glu Asn Ly - #s His Phe Cys Gly Gly               35         - #         40         - #         45                      - -  - - (2) INFORMATION FOR SEQ ID NO:66:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 390 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..390                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                              - - TTC GGC TTC AAG CTA AGT CAT TTG GTA AGT AA - #G TAC TGT GCT TGT GCA           48                                                                       Phe Gly Phe Lys Leu Ser His Leu Val Ser Ly - #s Tyr Cys Ala Cys Ala             1               5 - #                 10 - #                 15              - - TTA GCA TCG GCA CTG AAG TAC TCC ATC GAT CA - #T GGT CCT CGT ATC ATC           96                                                                       Leu Ala Ser Ala Leu Lys Tyr Ser Ile Asp Hi - #s Gly Pro Arg Ile Ile                        20     - #             25     - #             30                  - - GGA GGT GAA GTT GCA GGT GAA GGA TCA GCA CC - #T TAC CAG GTG TCC TTA          144                                                                       Gly Gly Glu Val Ala Gly Glu Gly Ser Ala Pr - #o Tyr Gln Val Ser Leu                    35         - #         40         - #         45                      - - AGA ACC AAG GAA GGA AAT CAT TTT TGC GGT GG - #A TCA ATA CTA AAT AAG          192                                                                       Arg Thr Lys Glu Gly Asn His Phe Cys Gly Gl - #y Ser Ile Leu Asn Lys                50             - #     55             - #     60                          - - CGA TGG GTT GTA ACT GCA GCA CAT TGT CTT GA - #A CCG GAA ATA TTA GAT          240                                                                       Arg Trp Val Val Thr Ala Ala His Cys Leu Gl - #u Pro Glu Ile Leu Asp            65                 - # 70                 - # 75                 - # 80       - - TCG GTA TAC GTC GGA TCC AAT CAC TTA GAC CG - #A AAA GGC AGA TAT TAC          288                                                                       Ser Val Tyr Val Gly Ser Asn His Leu Asp Ar - #g Lys Gly Arg Tyr Tyr                            85 - #                 90 - #                 95              - - GAC GTA GAA CGG TAT ATA ATT CAT GAA AAA TA - #T ATA GGA GAA CTA AAT          336                                                                       Asp Val Glu Arg Tyr Ile Ile His Glu Lys Ty - #r Ile Gly Glu Leu Asn                       100      - #           105      - #           110                  - - AAT TTT TAT GCT GAC ATC GGT CTA ATA AAA CT - #T GAT GGA AGA CTT AGA          384                                                                       Asn Phe Tyr Ala Asp Ile Gly Leu Ile Lys Le - #u Asp Gly Arg Leu Arg                   115          - #       120          - #       125                      - - ATT CAA                - #                  - #                  -      #          390                                                                  Ile Gln                                                                           130                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:67:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 130 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                              - - Phe Gly Phe Lys Leu Ser His Leu Val Ser Ly - #s Tyr Cys Ala Cys        Ala                                                                               1               5 - #                 10 - #                 15             - - Leu Ala Ser Ala Leu Lys Tyr Ser Ile Asp Hi - #s Gly Pro Arg Ile Ile                   20     - #             25     - #             30                  - - Gly Gly Glu Val Ala Gly Glu Gly Ser Ala Pr - #o Tyr Gln Val Ser Leu               35         - #         40         - #         45                      - - Arg Thr Lys Glu Gly Asn His Phe Cys Gly Gl - #y Ser Ile Leu Asn Lys           50             - #     55             - #     60                          - - Arg Trp Val Val Thr Ala Ala His Cys Leu Gl - #u Pro Glu Ile Leu Asp       65                 - # 70                 - # 75                 - # 80       - - Ser Val Tyr Val Gly Ser Asn His Leu Asp Ar - #g Lys Gly Arg Tyr Tyr                       85 - #                 90 - #                 95              - - Asp Val Glu Arg Tyr Ile Ile His Glu Lys Ty - #r Ile Gly Glu Leu Asn                  100      - #           105      - #           110                  - - Asn Phe Tyr Ala Asp Ile Gly Leu Ile Lys Le - #u Asp Gly Arg Leu Arg              115          - #       120          - #       125                      - - Ile Gln                                                                      130                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:68:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 240 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..240                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 218, change A to                      M.  At - # pos. aa 73, substitute Xaa."                         - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                              - - CGG GCT GCA GGA ATT CGG CAC GAG AAG AAA CT - #G CCA ATA TTA ATC GCC           48                                                                       Arg Ala Ala Gly Ile Arg His Glu Lys Lys Le - #u Pro Ile Leu Ile Ala             1               5 - #                 10 - #                 15              - - TTG ATC GGA TGC GTT CTT TCT GAA GAA ATA GA - #G GAT CGC ATT GTC GGC           96                                                                       Leu Ile Gly Cys Val Leu Ser Glu Glu Ile Gl - #u Asp Arg Ile Val Gly                        20     - #             25     - #             30                  - - GGA ACG GCA GTT GAT ATA AGA GGT TTT CCC TG - #G CAG GTA TCA ATT CAA          144                                                                       Gly Thr Ala Val Asp Ile Arg Gly Phe Pro Tr - #p Gln Val Ser Ile Gln                    35         - #         40         - #         45                      - - ACC GAA AAC CGT CAT TTT TGT GGT GGT TCT AT - #T ATC GAT AAA AGC TGG          192                                                                       Thr Glu Asn Arg His Phe Cys Gly Gly Ser Il - #e Ile Asp Lys Ser Trp                50             - #     55             - #     60                          - - ATA TTA ACT GCC GCA CAT TGT GTA CMC GAT AT - #G AAG ATG TCG AAC TGG          240                                                                       Ile Leu Thr Ala Ala His Cys Val Xaa Asp Me - #t Lys Met Ser Asn Trp            65                 - # 70                 - # 75                 - # 80       - -  - - (2) INFORMATION FOR SEQ ID NO:69:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 80 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                              - - Arg Ala Ala Gly Ile Arg His Glu Lys Lys Le - #u Pro Ile Leu Ile Ala        1               5 - #                 10 - #                 15              - - Leu Ile Gly Cys Val Leu Ser Glu Glu Ile Gl - #u Asp Arg Ile Val Gly                   20     - #             25     - #             30                  - - Gly Thr Ala Val Asp Ile Arg Gly Phe Pro Tr - #p Gln Val Ser Ile Gln               35         - #         40         - #         45                      - - Thr Glu Asn Arg His Phe Cys Gly Gly Ser Il - #e Ile Asp Lys Ser Trp           50             - #     55             - #     60                          - - Ile Leu Thr Ala Ala His Cys Val Xaa Asp Me - #t Lys Met Ser Asn Trp       65                 - # 70                 - # 75                 - # 80       - -  - - (2) INFORMATION FOR SEQ ID NO:70:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 177 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..177                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                              - - CAC GAG ATT TTA TTA AGC GCA TTA TTT GCA AG - #T GTA ATT TGC TCC TTT           48                                                                       His Glu Ile Leu Leu Ser Ala Leu Phe Ala Se - #r Val Ile Cys Ser Phe             1               5 - #                 10 - #                 15              - - AAC GCG GAA GTA CAA AAT CGA ATC GTT GGT GG - #C AAT GAT GTA AGT ATT           96                                                                       Asn Ala Glu Val Gln Asn Arg Ile Val Gly Gl - #y Asn Asp Val Ser Ile                        20     - #             25     - #             30                  - - TCA AAA ATT GGG TGG CAA GTA TCT ATT CAA AG - #T AAT AAA CAA CAT TTC          144                                                                       Ser Lys Ile Gly Trp Gln Val Ser Ile Gln Se - #r Asn Lys Gln His Phe                    35         - #         40         - #         45                      - - TGT GGT GGT TCA ATC ATT GCT AAA GAT GGG TC - #C                  -      #        177                                                                    Cys Gly Gly Ser Ile Ile Ala Lys Asp Gly Se - #r                                    50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:71:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 59 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                              - - His Glu Ile Leu Leu Ser Ala Leu Phe Ala Se - #r Val Ile Cys Ser        Phe                                                                               1               5 - #                 10 - #                 15             - - Asn Ala Glu Val Gln Asn Arg Ile Val Gly Gl - #y Asn Asp Val Ser Ile                   20     - #             25     - #             30                  - - Ser Lys Ile Gly Trp Gln Val Ser Ile Gln Se - #r Asn Lys Gln His Phe               35         - #         40         - #         45                      - - Cys Gly Gly Ser Ile Ile Ala Lys Asp Gly Se - #r                               50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:72:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 168 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..168                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                              - - ATC ATG GCA AAT TTT AGG CTA TTC ACC TTA CT - #A GCC TTG GTT TCA GTA           48                                                                       Ile Met Ala Asn Phe Arg Leu Phe Thr Leu Le - #u Ala Leu Val Ser Val             1               5 - #                 10 - #                 15              - - GCA ACT TCC AAA TAT ATT GAT CCA AGA ATA AT - #T GGA GGC GAA GAT GCT           96                                                                       Ala Thr Ser Lys Tyr Ile Asp Pro Arg Ile Il - #e Gly Gly Glu Asp Ala                        20     - #             25     - #             30                  - - CCT GAA GGC TCG GCT CCG TAC CAA GTT TCA TT - #G AGA AAT CAG AAT CTG          144                                                                       Pro Glu Gly Ser Ala Pro Tyr Gln Val Ser Le - #u Arg Asn Gln Asn Leu                    35         - #         40         - #         45                      - - GAG CAT TTC TGT GGT GGT TCC ATT     - #                  - #                   168                                                                     Glu His Phe Cys Gly Gly Ser Ile                                                    50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:73:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 56 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                              - - Ile Met Ala Asn Phe Arg Leu Phe Thr Leu Le - #u Ala Leu Val Ser Val        1               5 - #                 10 - #                 15              - - Ala Thr Ser Lys Tyr Ile Asp Pro Arg Ile Il - #e Gly Gly Glu Asp Ala                   20     - #             25     - #             30                  - - Pro Glu Gly Ser Ala Pro Tyr Gln Val Ser Le - #u Arg Asn Gln Asn Leu               35         - #         40         - #         45                      - - Glu His Phe Cys Gly Gly Ser Ile                                               50             - #     55                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:74:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 192 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..192                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                              - - GCA CGA GAT CGC ATT GTT GGA GGA TTG GAG GC - #G AAA AAT GGA TCA GCC           48                                                                       Ala Arg Asp Arg Ile Val Gly Gly Leu Glu Al - #a Lys Asn Gly Ser Ala             1               5 - #                 10 - #                 15              - - CCA TTC ATG GTT TCT TTG CAA GCG GAA GAC TA - #T TTT CAT TTT TGT GGA           96                                                                       Pro Phe Met Val Ser Leu Gln Ala Glu Asp Ty - #r Phe His Phe Cys Gly                        20     - #             25     - #             30                  - - TCC TCT ATT CTG AAT GAG AGA TGG GTT CTT AC - #T GCT GCT CAC TGT ATC          144                                                                       Ser Ser Ile Leu Asn Glu Arg Trp Val Leu Th - #r Ala Ala His Cys Ile                    35         - #         40         - #         45                      - - CAA CCA AAT GTA CAC AAG TAC GTT TAC GTC GG - #T TCG AAC AAC GTA GAA          192                                                                       Gln Pro Asn Val His Lys Tyr Val Tyr Val Gl - #y Ser Asn Asn Val Glu                50             - #     55             - #     60                          - -  - - (2) INFORMATION FOR SEQ ID NO:75:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 64 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                              - - Ala Arg Asp Arg Ile Val Gly Gly Leu Glu Al - #a Lys Asn Gly Ser Ala        1               5 - #                 10 - #                 15              - - Pro Phe Met Val Ser Leu Gln Ala Glu Asp Ty - #r Phe His Phe Cys Gly                   20     - #             25     - #             30                  - - Ser Ser Ile Leu Asn Glu Arg Trp Val Leu Th - #r Ala Ala His Cys Ile               35         - #         40         - #         45                      - - Gln Pro Asn Val His Lys Tyr Val Tyr Val Gl - #y Ser Asn Asn Val Glu           50             - #     55             - #     60                          - -  - - (2) INFORMATION FOR SEQ ID NO:76:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 207 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..204                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                              - - CCA ATC CAC GAT AGC CAA TAT GCA CTT TTG CA - #G ATA TGG GTC AAG GGT           48                                                                       Pro Ile His Asp Ser Gln Tyr Ala Leu Leu Gl - #n Ile Trp Val Lys Gly             1               5 - #                 10 - #                 15              - - GCA TGT AAG GGT GAT TCC GGT GGC CCC TTA GT - #C ATC AAT GGA CAA CTT           96                                                                       Ala Cys Lys Gly Asp Ser Gly Gly Pro Leu Va - #l Ile Asn Gly Gln Leu                        20     - #             25     - #             30                  - - CAT GGA ATT GTT TCC TGG GGC ATT CCT TGC GC - #T GTC GCA AGC CTG ATG          144                                                                       His Gly Ile Val Ser Trp Gly Ile Pro Cys Al - #a Val Ala Ser Leu Met                    35         - #         40         - #         45                      - - TAT TCA CAA GAG TTT CTC ATT ATG TCG ATT GG - #A TTA AAT CCA AAA TTG          192                                                                       Tyr Ser Gln Glu Phe Leu Ile Met Ser Ile Gl - #y Leu Asn Pro Lys Leu                50             - #     55             - #     60                          - - AAT AAA ATT GTT TAG           - #                  - #                      - #   207                                                                  Asn Lys Ile Val                                                                65                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:77:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 68 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                              - - Pro Ile His Asp Ser Gln Tyr Ala Leu Leu Gl - #n Ile Trp Val Lys Gly        1               5 - #                 10 - #                 15              - - Ala Cys Lys Gly Asp Ser Gly Gly Pro Leu Va - #l Ile Asn Gly Gln Leu                   20     - #             25     - #             30                  - - His Gly Ile Val Ser Trp Gly Ile Pro Cys Al - #a Val Ala Ser Leu Met               35         - #         40         - #         45                      - - Tyr Ser Gln Glu Phe Leu Ile Met Ser Ile Gl - #y Leu Asn Pro Lys Leu           50             - #     55             - #     60                          - - Asn Lys Ile Val                                                           65                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:78:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 159 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..156                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                              - - GGA GGT CCT TTG GCA ATC AAT GGT GAA CTT GT - #T GGT GTT ACT TCA TTC           48                                                                       Gly Gly Pro Leu Ala Ile Asn Gly Glu Leu Va - #l Gly Val Thr Ser Phe             1               5 - #                 10 - #                 15              - - ATT ATG GGG ACA TGT GGA GGA GGA CAT CCT GA - #T GTC TTC GGT CGA GTC           96                                                                       Ile Met Gly Thr Cys Gly Gly Gly His Pro As - #p Val Phe Gly Arg Val                        20     - #             25     - #             30                  - - CTT GAC TTC AAA CCA TGG ATT GAT TCT CAT AT - #G GCA AAT GAC GGC GCT          144                                                                       Leu Asp Phe Lys Pro Trp Ile Asp Ser His Me - #t Ala Asn Asp Gly Ala                    35         - #         40         - #         45                      - - AAT TCT TTT ATT TAA           - #                  - #                      - #   159                                                                  Asn Ser Phe Ile                                                                    50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:79:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 52 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                              - - Gly Gly Pro Leu Ala Ile Asn Gly Glu Leu Va - #l Gly Val Thr Ser Phe        1               5 - #                 10 - #                 15              - - Ile Met Gly Thr Cys Gly Gly Gly His Pro As - #p Val Phe Gly Arg Val                   20     - #             25     - #             30                  - - Leu Asp Phe Lys Pro Trp Ile Asp Ser His Me - #t Ala Asn Asp Gly Ala               35         - #         40         - #         45                      - - Asn Ser Phe Ile                                                               50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:80:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 779 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..699                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 371, change G to                      B.  At - # pos. aa 123, substitute Xaa."                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                              - - TT TTC TCA GCA CTC GTT GCC TTG TCT GCA GCT - # ATT CCT CAC TCC AAC            47                                                                          Phe Ser Ala Leu Val Ala Leu Ser Ala - #Ala Ile Pro His Ser Asn                  1             - #  5                - #  10                - #  15        - - AGA GTC GTT GGA GGA CTG GAA GCT GCA GAA GG - #T TCT GCA CCT TAC CAA           95                                                                       Arg Val Val Gly Gly Leu Glu Ala Ala Glu Gl - #y Ser Ala Pro Tyr Gln                            20 - #                 25 - #                 30              - - GTA TCC TTG CAA GTT GGT AAC TTC CAC TTC TG - #T GGT GGT TCA ATT TTG          143                                                                       Val Ser Leu Gln Val Gly Asn Phe His Phe Cy - #s Gly Gly Ser Ile Leu                        35     - #             40     - #             45                  - - AAC GAA TAT TGG GTT TTG ACT GCT GCT CAC TG - #T TTG GGT TAT GAC TTC          191                                                                       Asn Glu Tyr Trp Val Leu Thr Ala Ala His Cy - #s Leu Gly Tyr Asp Phe                    50         - #         55         - #         60                      - - GAC GTG GTA GTG GGA ACA AAC AAA CTT GAT CA - #A CCA GGT GAA AGA TAC          239                                                                       Asp Val Val Val Gly Thr Asn Lys Leu Asp Gl - #n Pro Gly Glu Arg Tyr                65             - #     70             - #     75                          - - CTC GTA GAA CAA ACT TTT GTT CAC CAA TTC GA - #C CAG GAA TCT TTA AGA          287                                                                       Leu Val Glu Gln Thr Phe Val His Gln Phe As - #p Gln Glu Ser Leu Arg            80                 - # 85                 - # 90                 - # 95       - - CAC GAT CTT GCT TTG GTA AAA GTG TCC AGC CC - #T ATC GAA TTC AAT GAT          335                                                                       His Asp Leu Ala Leu Val Lys Val Ser Ser Pr - #o Ile Glu Phe Asn Asp                           100  - #               105  - #               110              - - TAT GTT CAA CCA ATT CCA TTG GGC GAA ACT TA - #T GTB GGC GGT GAA GTT          383                                                                       Tyr Val Gln Pro Ile Pro Leu Gly Glu Thr Ty - #r Xaa Gly Gly Glu Val                       115      - #           120      - #           125                  - - GCT CGT CTT ACT GGA TGG GGA AGA CTT GGA GC - #T AAC TTG AAT GGA CCG          431                                                                       Ala Arg Leu Thr Gly Trp Gly Arg Leu Gly Al - #a Asn Leu Asn Gly Pro                   130          - #       135          - #       140                      - - AAT GAA CTC CAA GAA CTT AAC ACT GTC ACA TT - #A AGC CAC CAG CAA TGT          479                                                                       Asn Glu Leu Gln Glu Leu Asn Thr Val Thr Le - #u Ser His Gln Gln Cys               145              - #   150              - #   155                          - - GTA AGA CAA CAA ATT TAT CCA GTA TAC GAC AG - #C CAA CTT TGC ACA TTT          527                                                                       Val Arg Gln Gln Ile Tyr Pro Val Tyr Asp Se - #r Gln Leu Cys Thr Phe           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - GTT GGC AGT GGA CGA GGC GCC TGC AAC GGT GA - #C TCT GGT GGT CCA        TTG      575                                                                    Val Gly Ser Gly Arg Gly Ala Cys Asn Gly As - #p Ser Gly Gly Pro Leu                          180  - #               185  - #               190              - - GTC GTC AAT GGA GAA CTT CAC GGA GTC GTC TC - #C TGG GGA ATC CCC TGC          623                                                                       Val Val Asn Gly Glu Leu His Gly Val Val Se - #r Trp Gly Ile Pro Cys                       195      - #           200      - #           205                  - - GCC GTT GGA TTA CCC GAT GTC TTC ACA AGA GT - #T TCA CAC TAC GCT GAC          671                                                                       Ala Val Gly Leu Pro Asp Val Phe Thr Arg Va - #l Ser His Tyr Ala Asp                   210          - #       215          - #       220                      - - TGG ATT AGA GAG ACC ATG GAA AAT AAC T AATT - #TTTAAT GGCATATTAT              719                                                                       Trp Ile Arg Glu Thr Met Glu Asn Asn                                               225              - #   230                                                 - - TGTATTGTCT GTGATGAAAA TTAATAAAAA CGTGATAGAT TAAAAAAAAA AA -             #AAAAAAAA    779                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:81:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 232 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                              - - Phe Ser Ala Leu Val Ala Leu Ser Ala Ala Il - #e Pro His Ser Asn        Arg                                                                               1               5 - #                 10 - #                 15             - - Val Val Gly Gly Leu Glu Ala Ala Glu Gly Se - #r Ala Pro Tyr Gln Val                   20     - #             25     - #             30                  - - Ser Leu Gln Val Gly Asn Phe His Phe Cys Gl - #y Gly Ser Ile Leu Asn               35         - #         40         - #         45                      - - Glu Tyr Trp Val Leu Thr Ala Ala His Cys Le - #u Gly Tyr Asp Phe Asp           50             - #     55             - #     60                          - - Val Val Val Gly Thr Asn Lys Leu Asp Gln Pr - #o Gly Glu Arg Tyr Leu       65                 - # 70                 - # 75                 - # 80       - - Val Glu Gln Thr Phe Val His Gln Phe Asp Gl - #n Glu Ser Leu Arg His                       85 - #                 90 - #                 95              - - Asp Leu Ala Leu Val Lys Val Ser Ser Pro Il - #e Glu Phe Asn Asp Tyr                  100      - #           105      - #           110                  - - Val Gln Pro Ile Pro Leu Gly Glu Thr Tyr Xa - #a Gly Gly Glu Val Ala              115          - #       120          - #       125                      - - Arg Leu Thr Gly Trp Gly Arg Leu Gly Ala As - #n Leu Asn Gly Pro Asn          130              - #   135              - #   140                          - - Glu Leu Gln Glu Leu Asn Thr Val Thr Leu Se - #r His Gln Gln Cys Val      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Arg Gln Gln Ile Tyr Pro Val Tyr Asp Ser Gl - #n Leu Cys Thr Phe        Val                                                                                             165  - #               170  - #               175             - - Gly Ser Gly Arg Gly Ala Cys Asn Gly Asp Se - #r Gly Gly Pro Leu Val                  180      - #           185      - #           190                  - - Val Asn Gly Glu Leu His Gly Val Val Ser Tr - #p Gly Ile Pro Cys Ala              195          - #       200          - #       205                      - - Val Gly Leu Pro Asp Val Phe Thr Arg Val Se - #r His Tyr Ala Asp Trp          210              - #   215              - #   220                          - - Ile Arg Glu Thr Met Glu Asn Asn                                          225                 2 - #30                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:82:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 944 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..768                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                              - - CT GAA GTG TGG ATT CAT CGT TGT TTT CCT TGT - # GGC GGC AGC TTG GGT            47                                                                        - -    Glu Val Trp Ile His Arg Cys Phe Pro - #Cys Gly Gly Ser Leu Gly             1             - #  5                - #  10                - #  15        - - GAA GAT TCG GTC GTC GAC CGC ATC GTC GGC GG - #C ACC AGT GTT AAA ATT           95                                                                       Glu Asp Ser Val Val Asp Arg Ile Val Gly Gl - #y Thr Ser Val Lys Ile                            20 - #                 25 - #                 30              - - GAG AAC TTC GGA TGG CAA GTG TCC TTA TTC GA - #T CGT AAG GGT CAC TTT          143                                                                       Glu Asn Phe Gly Trp Gln Val Ser Leu Phe As - #p Arg Lys Gly His Phe                        35     - #             40     - #             45                  - - TGC GGT GGT TCT ATA ATC AGC GAC GAA TGG GT - #C TTG ACT GCT GCA CAT          191                                                                       Cys Gly Gly Ser Ile Ile Ser Asp Glu Trp Va - #l Leu Thr Ala Ala His                    50         - #         55         - #         60                      - - TGC GTA TAC GAT TAT TTC TCG CCA AAG CAA TA - #T GGA GTG CGT GTC GGA          239                                                                       Cys Val Tyr Asp Tyr Phe Ser Pro Lys Gln Ty - #r Gly Val Arg Val Gly                65             - #     70             - #     75                          - - AGC AGT TTA CGC AAC AAA GGT GGA GTC CTT CA - #C AGA ATT TCC AGG GTA          287                                                                       Ser Ser Leu Arg Asn Lys Gly Gly Val Leu Hi - #s Arg Ile Ser Arg Val            80                 - # 85                 - # 90                 - # 95       - - CAC ATT CAC CCA GAC TAC GAC ACG GTC AGC TA - #C GAC AAT GAC GTC GCG          335                                                                       His Ile His Pro Asp Tyr Asp Thr Val Ser Ty - #r Asp Asn Asp Val Ala                           100  - #               105  - #               110              - - CTC CTG AAA GTT GAA ACC AAA TTT AAA CTA AA - #C GGC AGG AGC GTT CGC          383                                                                       Leu Leu Lys Val Glu Thr Lys Phe Lys Leu As - #n Gly Arg Ser Val Arg                       115      - #           120      - #           125                  - - AAA GTT AAA TTG GTT GAC GAA GAT CAC GAG GT - #T GAT GAT GGT GCC CGG          431                                                                       Lys Val Lys Leu Val Asp Glu Asp His Glu Va - #l Asp Asp Gly Ala Arg                   130          - #       135          - #       140                      - - CTC ACC GTC ACT GGA TGG GGC AAA TTA AGT GA - #A TCA GGA CCC AAG CCA          479                                                                       Leu Thr Val Thr Gly Trp Gly Lys Leu Ser Gl - #u Ser Gly Pro Lys Pro               145              - #   150              - #   155                          - - GTA AAT CTA CAA GGA GTA AAA GTG CCT TAT GT - #G GAC CAA GAT ACA TGC          527                                                                       Val Asn Leu Gln Gly Val Lys Val Pro Tyr Va - #l Asp Gln Asp Thr Cys           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - TCT GAC AGC TAC GTC TTT GCA GGA AAA GAT AT - #C ACC GAA AAC ATG        TTG      575                                                                    Ser Asp Ser Tyr Val Phe Ala Gly Lys Asp Il - #e Thr Glu Asn Met Leu                          180  - #               185  - #               190              - - TGT GCC GGA GTT AGA AGA GGT GGC AAG GAC TC - #C TGC CAG GGT GAC AGC          623                                                                       Cys Ala Gly Val Arg Arg Gly Gly Lys Asp Se - #r Cys Gln Gly Asp Ser                       195      - #           200      - #           205                  - - GGT GGT CCA CTT GTG GAC GAA AAC AAA AAT CT - #G GTC GGA GTC GTC TCT          671                                                                       Gly Gly Pro Leu Val Asp Glu Asn Lys Asn Le - #u Val Gly Val Val Ser                   210          - #       215          - #       220                      - - TGG GGA AAT GGT TGT GCC AGA CCA AAC ATG CC - #A GGA GTA TAC GCT AAA          719                                                                       Trp Gly Asn Gly Cys Ala Arg Pro Asn Met Pr - #o Gly Val Tyr Ala Lys               225              - #   230              - #   235                          - - GTT GCT GCT TCT AGC ATT AGA GAG TTC ATT CG - #C AAA AAA ACT GGT CTT     T    768                                                                       Val Ala Ala Ser Ser Ile Arg Glu Phe Ile Ar - #g Lys Lys Thr Gly Leu           240                 2 - #45                 2 - #50                 2 -      #55                                                                              - - AATTTCCTTA TATGAACAAA TGTTCCACCA AAAATATAGT TTAGATTTTA GT -            #ATAATAAA    828                                                                 - - TCCTTTGTGA TTCATGCAAA TATTTTGTTT TATTTATTTA TTTACTTTAT TC -            #AAACGAAT    888                                                                 - - GTATAAAGTG AATTAACAAT AAAAATGTTA GTGTTGCCAA AAAAAAAAAA AA - #AAAA            944                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:83:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 255 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                              - - Glu Val Trp Ile His Arg Cys Phe Pro Cys Gl - #y Gly Ser Leu Gly Glu        1               5 - #                 10 - #                 15              - - Asp Ser Val Val Asp Arg Ile Val Gly Gly Th - #r Ser Val Lys Ile Glu                   20     - #             25     - #             30                  - - Asn Phe Gly Trp Gln Val Ser Leu Phe Asp Ar - #g Lys Gly His Phe Cys               35         - #         40         - #         45                      - - Gly Gly Ser Ile Ile Ser Asp Glu Trp Val Le - #u Thr Ala Ala His Cys           50             - #     55             - #     60                          - - Val Tyr Asp Tyr Phe Ser Pro Lys Gln Tyr Gl - #y Val Arg Val Gly Ser       65                 - # 70                 - # 75                 - # 80       - - Ser Leu Arg Asn Lys Gly Gly Val Leu His Ar - #g Ile Ser Arg Val His                       85 - #                 90 - #                 95              - - Ile His Pro Asp Tyr Asp Thr Val Ser Tyr As - #p Asn Asp Val Ala Leu                  100      - #           105      - #           110                  - - Leu Lys Val Glu Thr Lys Phe Lys Leu Asn Gl - #y Arg Ser Val Arg Lys              115          - #       120          - #       125                      - - Val Lys Leu Val Asp Glu Asp His Glu Val As - #p Asp Gly Ala Arg Leu          130              - #   135              - #   140                          - - Thr Val Thr Gly Trp Gly Lys Leu Ser Glu Se - #r Gly Pro Lys Pro Val      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Asn Leu Gln Gly Val Lys Val Pro Tyr Val As - #p Gln Asp Thr Cys        Ser                                                                                             165  - #               170  - #               175             - - Asp Ser Tyr Val Phe Ala Gly Lys Asp Ile Th - #r Glu Asn Met Leu Cys                  180      - #           185      - #           190                  - - Ala Gly Val Arg Arg Gly Gly Lys Asp Ser Cy - #s Gln Gly Asp Ser Gly              195          - #       200          - #       205                      - - Gly Pro Leu Val Asp Glu Asn Lys Asn Leu Va - #l Gly Val Val Ser Trp          210              - #   215              - #   220                          - - Gly Asn Gly Cys Ala Arg Pro Asn Met Pro Gl - #y Val Tyr Ala Lys Val      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Ala Ala Ser Ser Ile Arg Glu Phe Ile Arg Ly - #s Lys Thr Gly Leu                         245  - #               250  - #               255              - -  - - (2) INFORMATION FOR SEQ ID NO:84:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 157 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..157                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                              - - GGA ATT CGG CAC GAG TAT CTG TGC CCT CGC AG - #T CTG CAC CCT TGG CGT           48                                                                       Gly Ile Arg His Glu Tyr Leu Cys Pro Arg Se - #r Leu His Pro Trp Arg             1               5 - #                 10 - #                 15              - - AGC GTT CCT GAC TTT TGG AAC AGG TTA GAT GG - #C AGA ATC GTT GGA GGA           96                                                                       Ser Val Pro Asp Phe Trp Asn Arg Leu Asp Gl - #y Arg Ile Val Gly Gly                        20     - #             25     - #             30                  - - CAC GAT ACT AGC ATT GAT AAC ATC CTC ATG CA - #A GTA TCT TTG AGT TTA          144                                                                       His Asp Thr Ser Ile Asp Asn Ile Leu Met Gl - #n Val Ser Leu Ser Leu                    35         - #         40         - #         45                      - - CAC AAA CCA CAA  T          - #                  - #                      - #     157                                                                  His Lys Pro Gln                                                                    50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:85:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 52 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                              - - Gly Ile Arg His Glu Tyr Leu Cys Pro Arg Se - #r Leu His Pro Trp Arg        1               5 - #                 10 - #                 15              - - Ser Val Pro Asp Phe Trp Asn Arg Leu Asp Gl - #y Arg Ile Val Gly Gly                   20     - #             25     - #             30                  - - His Asp Thr Ser Ile Asp Asn Ile Leu Met Gl - #n Val Ser Leu Ser Leu               35         - #         40         - #         45                      - - His Lys Pro Gln                                                               50                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:86:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 218 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..218                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                              - - AA GAT GTA GAA TTA ACA CCT GGA ACT ATG TGC - # ACT GTT ACT GGA TGG            47                                                                          Asp Val Glu Leu Thr Pro Gly Thr Met - #Cys Thr Val Thr Gly Trp                  1             - #  5                - #  10                - #  15        - - GGA TCA ACT GGA TCT GGT GGT CCA ATT ACA AA - #T GTT CTA CAA GAA GTC           95                                                                       Gly Ser Thr Gly Ser Gly Gly Pro Ile Thr As - #n Val Leu Gln Glu Val                            20 - #                 25 - #                 30              - - GAA GTT CCA TTT ATC GAC TTC AAC ACC TGC CG - #A AAA TCC TAC TCA ACC          143                                                                       Glu Val Pro Phe Ile Asp Phe Asn Thr Cys Ar - #g Lys Ser Tyr Ser Thr                        35     - #             40     - #             45                  - - AGC TTA ACC GAC CGT ATG TTC TGC GCT GGA TT - #T TTG GGA ATT GGT GGT          191                                                                       Ser Leu Thr Asp Arg Met Phe Cys Ala Gly Ph - #e Leu Gly Ile Gly Gly                    50         - #         55         - #         60                      - - AAG GAC GCT TGC CAA GGC GAC TCC GGA    - #                  - #                218                                                                     Lys Asp Ala Cys Gln Gly Asp Ser Gly                                                65             - #     70                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:87:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 72 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                              - - Asp Val Glu Leu Thr Pro Gly Thr Met Cys Th - #r Val Thr Gly Trp Gly        1               5 - #                 10 - #                 15              - - Ser Thr Gly Ser Gly Gly Pro Ile Thr Asn Va - #l Leu Gln Glu Val Glu                   20     - #             25     - #             30                  - - Val Pro Phe Ile Asp Phe Asn Thr Cys Arg Ly - #s Ser Tyr Ser Thr Ser               35         - #         40         - #         45                      - - Leu Thr Asp Arg Met Phe Cys Ala Gly Phe Le - #u Gly Ile Gly Gly Lys           50             - #     55             - #     60                          - - Asp Ala Cys Gln Gly Asp Ser Gly                                           65                 - # 70                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:88:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 932 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..770                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 276, change G to                      K.  At - # pos. bp 332, change G to K.  At pos. - #aa 92      and                                                                                            aa 111, - #substitute Xaa."                                     - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                              - - G TTC ATA TTT GTG CTC GTT TGC GTT GGA TTG - #AGC GCC GTC TCA TCT             46                                                                         Phe Ile Phe Val Leu Val Cys Val Gly L - #eu Ser Ala Val Ser Ser                 1              - # 5                 - # 10                 - # 15         - - TAC AAG ATA AAG GAT GGA TTA GAT GGG CGC AT - #T GTT GGA GGA CAA GAT           94                                                                       Tyr Lys Ile Lys Asp Gly Leu Asp Gly Arg Il - #e Val Gly Gly Gln Asp                            20 - #                 25 - #                 30              - - GCT GAT ATT GCC AAA TAT GGC TAT CAA GCT TC - #A CTC CAA GTA TTT AAC          142                                                                       Ala Asp Ile Ala Lys Tyr Gly Tyr Gln Ala Se - #r Leu Gln Val Phe Asn                        35     - #             40     - #             45                  - - GAA CAT TTC TGT GGA GCT TCA ATA TTG AAT AA - #T TAT TGG ATT GTC ACA          190                                                                       Glu His Phe Cys Gly Ala Ser Ile Leu Asn As - #n Tyr Trp Ile Val Thr                    50         - #         55         - #         60                      - - GCA GCT CAT TGC ATA TAT GAT GAA TTC ACG TA - #T TCA GTT CGA GTC GGC          238                                                                       Ala Ala His Cys Ile Tyr Asp Glu Phe Thr Ty - #r Ser Val Arg Val Gly                65             - #     70             - #     75                          - - ACC AGT TTC CAA GGA AGA CGT GGT TCC GTT CA - #T CCT GKG GCA CAA ATT          286                                                                       Thr Ser Phe Gln Gly Arg Arg Gly Ser Val Hi - #s Pro Xaa Ala Gln Ile            80                 - # 85                 - # 90                 - # 95       - - ATC AAG CAT CCT GCA TAC GGT AAT GTA ACT GA - #C ATC GAT ATG GAA KGC          334                                                                       Ile Lys His Pro Ala Tyr Gly Asn Val Thr As - #p Ile Asp Met Glu Xaa                           100  - #               105  - #               110              - - GCC CTC ATC AAG GTT CGA AGA CCA TTC CGG TT - #G AAT AAC AGA ACT GTT          382                                                                       Ala Leu Ile Lys Val Arg Arg Pro Phe Arg Le - #u Asn Asn Arg Thr Val                       115      - #           120      - #           125                  - - AGA ACA GTC AAA CTT ACT GAT GTT GGA AAA GA - #C ATG CCA TCA GGA GAA          430                                                                       Arg Thr Val Lys Leu Thr Asp Val Gly Lys As - #p Met Pro Ser Gly Glu                   130          - #       135          - #       140                      - - TTA GCC ACT GTT ACT GGC TGG GGA AAT TTA GG - #G GAA GAT GAA GAC GAC          478                                                                       Leu Ala Thr Val Thr Gly Trp Gly Asn Leu Gl - #y Glu Asp Glu Asp Asp               145              - #   150              - #   155                          - - CCC GAA CAA CTG CAA TAT GTA AAG GTA CCT AT - #T GTT AAC TGG ACT CAG          526                                                                       Pro Glu Gln Leu Gln Tyr Val Lys Val Pro Il - #e Val Asn Trp Thr Gln           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - TGC AAA ACT ATA TAT GGA AAT GAA GGA CTA AT - #A ATT ACC CAA AAT        ATG      574                                                                    Cys Lys Thr Ile Tyr Gly Asn Glu Gly Leu Il - #e Ile Thr Gln Asn Met                          180  - #               185  - #               190              - - ATT TGT GCT GGT TAT CCT GAA GGC GGT AAG GA - #C TCT TGC CAA GGA GAT          622                                                                       Ile Cys Ala Gly Tyr Pro Glu Gly Gly Lys As - #p Ser Cys Gln Gly Asp                       195      - #           200      - #           205                  - - AGC GGT GGC CCA CTC GTC AAC TCT AAG GGA GT - #T CTG CAT GGA ATA GTG          670                                                                       Ser Gly Gly Pro Leu Val Asn Ser Lys Gly Va - #l Leu His Gly Ile Val                   210          - #       215          - #       220                      - - TCT TGG GGA ATA GGA TGT GCA CGA CCC GAA AT - #C CCA GGA GTA TAT ACC          718                                                                       Ser Trp Gly Ile Gly Cys Ala Arg Pro Glu Il - #e Pro Gly Val Tyr Thr               225              - #   230              - #   235                          - - CGA GTG GCT TCA AAA CCA ATA AGA GAA TTT AT - #C AAA ATG CAC ACT GGA          766                                                                       Arg Val Ala Ser Lys Pro Ile Arg Glu Phe Il - #e Lys Met His Thr Gly           240                 2 - #45                 2 - #50                 2 -      #55                                                                              - - ATA T AAGAGTTTTA ACTTATAATA TTACAAATAT TTTTTGATAT TCCT - #TAATTT             820                                                                      Ile                                                                            - - CAATGATATA CTAAGACGAG ATGTTTTACA AAATTTTGAT ACTCAACTAA CA -             #AATTAAAC    880                                                                 - - CATATTACTA CTCAAATAAA TATCACTAAT AATCAAAAAA AAAAAAAAAA AA - #                932                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:89:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 256 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                              - - Phe Ile Phe Val Leu Val Cys Val Gly Leu Se - #r Ala Val Ser Ser Tyr        1               5 - #                 10 - #                 15              - - Lys Ile Lys Asp Gly Leu Asp Gly Arg Ile Va - #l Gly Gly Gln Asp Ala                   20     - #             25     - #             30                  - - Asp Ile Ala Lys Tyr Gly Tyr Gln Ala Ser Le - #u Gln Val Phe Asn Glu               35         - #         40         - #         45                      - - His Phe Cys Gly Ala Ser Ile Leu Asn Asn Ty - #r Trp Ile Val Thr Ala           50             - #     55             - #     60                          - - Ala His Cys Ile Tyr Asp Glu Phe Thr Tyr Se - #r Val Arg Val Gly Thr       65                 - # 70                 - # 75                 - # 80       - - Ser Phe Gln Gly Arg Arg Gly Ser Val His Pr - #o Xaa Ala Gln Ile Ile                       85 - #                 90 - #                 95              - - Lys His Pro Ala Tyr Gly Asn Val Thr Asp Il - #e Asp Met Glu Xaa Ala                  100      - #           105      - #           110                  - - Leu Ile Lys Val Arg Arg Pro Phe Arg Leu As - #n Asn Arg Thr Val Arg              115          - #       120          - #       125                      - - Thr Val Lys Leu Thr Asp Val Gly Lys Asp Me - #t Pro Ser Gly Glu Leu          130              - #   135              - #   140                          - - Ala Thr Val Thr Gly Trp Gly Asn Leu Gly Gl - #u Asp Glu Asp Asp Pro      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Glu Gln Leu Gln Tyr Val Lys Val Pro Ile Va - #l Asn Trp Thr Gln        Cys                                                                                             165  - #               170  - #               175             - - Lys Thr Ile Tyr Gly Asn Glu Gly Leu Ile Il - #e Thr Gln Asn Met Ile                  180      - #           185      - #           190                  - - Cys Ala Gly Tyr Pro Glu Gly Gly Lys Asp Se - #r Cys Gln Gly Asp Ser              195          - #       200          - #       205                      - - Gly Gly Pro Leu Val Asn Ser Lys Gly Val Le - #u His Gly Ile Val Ser          210              - #   215              - #   220                          - - Trp Gly Ile Gly Cys Ala Arg Pro Glu Ile Pr - #o Gly Val Tyr Thr Arg      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Val Ala Ser Lys Pro Ile Arg Glu Phe Ile Ly - #s Met His Thr Gly        Ile                                                                                             245  - #               250  - #               255             - -  - - (2) INFORMATION FOR SEQ ID NO:90:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 894 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..766                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 560, change G to                      N.  At - # pos. aa 187, substitute Xaa."                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                              - - CCG GCG GTG ATT GTG TCA TTT GTT CTG GCT TG - #T GCA TTT TCT GTA CAG           48                                                                       Pro Ala Val Ile Val Ser Phe Val Leu Ala Cy - #s Ala Phe Ser Val Gln             1               5 - #                 10 - #                 15              - - GCT CTT CCA TCA AGC AGA ATT GTC AAT GGA CT - #T GAA GCA GGA GTT GGA           96                                                                       Ala Leu Pro Ser Ser Arg Ile Val Asn Gly Le - #u Glu Ala Gly Val Gly                        20     - #             25     - #             30                  - - CAA TTT CCA ATT CAG GTT TTC TTA GAC TTG AC - #A AAT ATC AGA GAC GAA          144                                                                       Gln Phe Pro Ile Gln Val Phe Leu Asp Leu Th - #r Asn Ile Arg Asp Glu                    35         - #         40         - #         45                      - - AAA TCC AGA TGT GGT GGT GCT TTG TTA TCA GA - #T TCA TGG GTT TTG ACT          192                                                                       Lys Ser Arg Cys Gly Gly Ala Leu Leu Ser As - #p Ser Trp Val Leu Thr                50             - #     55             - #     60                          - - GCT GCT CAT TGT TTT GAT GAT TTG AAG TCT AT - #G GTA GTG TCC GTT GGT          240                                                                       Ala Ala His Cys Phe Asp Asp Leu Lys Ser Me - #t Val Val Ser Val Gly            65                 - # 70                 - # 75                 - # 80       - - GCT CAT GAT GTC AGC AAA TCT GAA GAA CCT CA - #C AGG CAA ACC AGG AAA          288                                                                       Ala His Asp Val Ser Lys Ser Glu Glu Pro Hi - #s Arg Gln Thr Arg Lys                            85 - #                 90 - #                 95              - - CCT GAA AGG TAC TTC CAG CAT GAA AAA TAC GA - #C AGG GCA AAT CTT GCA          336                                                                       Pro Glu Arg Tyr Phe Gln His Glu Lys Tyr As - #p Arg Ala Asn Leu Ala                       100      - #           105      - #           110                  - - TAT GAT CTT GGT TTG TTG AAA TTG GAC AAA CC - #A GTG GAA TTG AAT GAT          384                                                                       Tyr Asp Leu Gly Leu Leu Lys Leu Asp Lys Pr - #o Val Glu Leu Asn Asp                   115          - #       120          - #       125                      - - TTC GTG AAA CTC ACA AAA TTG AAC AAA GAC AA - #A ACT GAA ACT TTT GTC          432                                                                       Phe Val Lys Leu Thr Lys Leu Asn Lys Asp Ly - #s Thr Glu Thr Phe Val               130              - #   135              - #   140                          - - GGC AAA ACT GCA ACT GTT AGT GGA TGG GCA TC - #T CCA AAG ATT TCC CCT          480                                                                       Gly Lys Thr Ala Thr Val Ser Gly Trp Ala Se - #r Pro Lys Ile Ser Pro           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - GCT TTC GAA TTG CCT GAC AAA CTA CAG TAC AC - #A ACT TTG GAA GTC        CAA      528                                                                    Ala Phe Glu Leu Pro Asp Lys Leu Gln Tyr Th - #r Thr Leu Glu Val Gln                          165  - #               170  - #               175              - - CCA AGT GAA GAC TGC AAA AAA GTA TGG GCC CN - #T TAC ATG CGC GAC TAC          576                                                                       Pro Ser Glu Asp Cys Lys Lys Val Trp Ala Xa - #a Tyr Met Arg Asp Tyr                       180      - #           185      - #           190                  - - ATC CTT TGT GCC AAA TTT GAA AAA CAA AAC AT - #T TGC ACT GGT GAC AGT          624                                                                       Ile Leu Cys Ala Lys Phe Glu Lys Gln Asn Il - #e Cys Thr Gly Asp Ser                   195          - #       200          - #       205                      - - GGC GGT CCA TTG ACC ATT GAT GGT GTC CAA GT - #T GGT GTG GTG AGT TTT          672                                                                       Gly Gly Pro Leu Thr Ile Asp Gly Val Gln Va - #l Gly Val Val Ser Phe               210              - #   215              - #   220                          - - GGA AGT GTT CCT TGT GCC AGA GGA AAT CCT TC - #A GGA TTT ACC AAT GTT          720                                                                       Gly Ser Val Pro Cys Ala Arg Gly Asn Pro Se - #r Gly Phe Thr Asn Val           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - GCT CAT TTT GTG GAT TGG ATT CAA GAA CAT AC - #T GGA TTG GAA TTG T            766                                                                      Ala His Phe Val Asp Trp Ile Gln Glu His Th - #r Gly Leu Glu Leu                               245  - #               250  - #               255              - - AAAAATAAAA CTCAAACTAA AAAAAAAATA ATTTAATTGC ACTGAAAAAT TT -             #TTCAAGAA    826                                                                 - - AAGTTTGGAT CGTTTTGTAA TTGAATGACA ATAAAAGCGT AAATTAGAAA AA -            #AAAAAAAA    886                                                                 - - AAAAACTC                - #                  - #                       - #         894                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:91:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 255 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                              - - Pro Ala Val Ile Val Ser Phe Val Leu Ala Cy - #s Ala Phe Ser Val        Gln                                                                               1               5 - #                 10 - #                 15             - - Ala Leu Pro Ser Ser Arg Ile Val Asn Gly Le - #u Glu Ala Gly Val Gly                   20     - #             25     - #             30                  - - Gln Phe Pro Ile Gln Val Phe Leu Asp Leu Th - #r Asn Ile Arg Asp Glu               35         - #         40         - #         45                      - - Lys Ser Arg Cys Gly Gly Ala Leu Leu Ser As - #p Ser Trp Val Leu Thr           50             - #     55             - #     60                          - - Ala Ala His Cys Phe Asp Asp Leu Lys Ser Me - #t Val Val Ser Val Gly       65                 - # 70                 - # 75                 - # 80       - - Ala His Asp Val Ser Lys Ser Glu Glu Pro Hi - #s Arg Gln Thr Arg Lys                       85 - #                 90 - #                 95              - - Pro Glu Arg Tyr Phe Gln His Glu Lys Tyr As - #p Arg Ala Asn Leu Ala                  100      - #           105      - #           110                  - - Tyr Asp Leu Gly Leu Leu Lys Leu Asp Lys Pr - #o Val Glu Leu Asn Asp              115          - #       120          - #       125                      - - Phe Val Lys Leu Thr Lys Leu Asn Lys Asp Ly - #s Thr Glu Thr Phe Val          130              - #   135              - #   140                          - - Gly Lys Thr Ala Thr Val Ser Gly Trp Ala Se - #r Pro Lys Ile Ser Pro      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ala Phe Glu Leu Pro Asp Lys Leu Gln Tyr Th - #r Thr Leu Glu Val        Gln                                                                                             165  - #               170  - #               175             - - Pro Ser Glu Asp Cys Lys Lys Val Trp Ala Xa - #a Tyr Met Arg Asp Tyr                  180      - #           185      - #           190                  - - Ile Leu Cys Ala Lys Phe Glu Lys Gln Asn Il - #e Cys Thr Gly Asp Ser              195          - #       200          - #       205                      - - Gly Gly Pro Leu Thr Ile Asp Gly Val Gln Va - #l Gly Val Val Ser Phe          210              - #   215              - #   220                          - - Gly Ser Val Pro Cys Ala Arg Gly Asn Pro Se - #r Gly Phe Thr Asn Val      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Ala His Phe Val Asp Trp Ile Gln Glu His Th - #r Gly Leu Glu Leu                         245  - #               250  - #               255              - -  - - (2) INFORMATION FOR SEQ ID NO:92:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 299 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..299                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 178/179, change                       G/C to - #V/Y.  At pos. aa 59, substitue Xaa."                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                              - - AA TTC GGC ACG AGA GTT AGT CTT TCC AAT TCG - # ATC AGA CCT TCT TGT            47                                                                        - -    Phe Gly Thr Arg Val Ser Leu Ser Asn - #Ser Ile Arg Pro Ser Cys             1             - #  5                - #  10                - #  15        - - TTA TGG GCC AAT GAC GAG TTC GAC ACA GAT AG - #T TCA ATT GCT ACT GGT           95                                                                       Leu Trp Ala Asn Asp Glu Phe Asp Thr Asp Se - #r Ser Ile Ala Thr Gly                            20 - #                 25 - #                 30              - - TGG GGA AAG ATA GAC TAT GCT GAG AGC AGA AG - #T GAT GAC CTA CTG AAA          143                                                                       Trp Gly Lys Ile Asp Tyr Ala Glu Ser Arg Se - #r Asp Asp Leu Leu Lys                        35     - #             40     - #             45                  - - GTA GTA CTG AAA ATT ATT GAT AAT AGG CAA TG - #C GVY CCC TTA TAC GTT          191                                                                       Val Val Leu Lys Ile Ile Asp Asn Arg Gln Cy - #s Xaa Pro Leu Tyr Val                    50         - #         55         - #         60                      - - GAT CAG ATT AAT AGA AGA AGA TTG AGA AAT GG - #A ATT GTA GAT ACA CAG          239                                                                       Asp Gln Ile Asn Arg Arg Arg Leu Arg Asn Gl - #y Ile Val Asp Thr Gln                65             - #     70             - #     75                          - - ATG TGT GCA GGA GAA TTG GAT GGT GGC AAA GA - #C ACT TGC CAG GGA GAT          287                                                                       Met Cys Ala Gly Glu Leu Asp Gly Gly Lys As - #p Thr Cys Gln Gly Asp            80                 - # 85                 - # 90                 - # 95       - - TCA GGT GGT CCT            - #                  - #                      - #      299                                                                  Ser Gly Gly Pro                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:93:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 99 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                              - - Phe Gly Thr Arg Val Ser Leu Ser Asn Ser Il - #e Arg Pro Ser Cys Leu        1               5 - #                 10 - #                 15              - - Trp Ala Asn Asp Glu Phe Asp Thr Asp Ser Se - #r Ile Ala Thr Gly Trp                   20     - #             25     - #             30                  - - Gly Lys Ile Asp Tyr Ala Glu Ser Arg Ser As - #p Asp Leu Leu Lys Val               35         - #         40         - #         45                      - - Val Leu Lys Ile Ile Asp Asn Arg Gln Cys Xa - #a Pro Leu Tyr Val Asp           50             - #     55             - #     60                          - - Gln Ile Asn Arg Arg Arg Leu Arg Asn Gly Il - #e Val Asp Thr Gln Met       65                 - # 70                 - # 75                 - # 80       - - Cys Ala Gly Glu Leu Asp Gly Gly Lys Asp Th - #r Cys Gln Gly Asp Ser                       85 - #                 90 - #                 95              - - Gly Gly Pro                                                               - -  - - (2) INFORMATION FOR SEQ ID NO:94:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 266 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..266                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                              - - CTT GCA TAT CCG CTC AAG TTT AGC GAA GAC AT - #C CAA CCG ATC ATG ATG           48                                                                       Leu Ala Tyr Pro Leu Lys Phe Ser Glu Asp Il - #e Gln Pro Ile Met Met             1               5 - #                 10 - #                 15              - - GCC GAA AAG GAC TAC GAA CCA CCA GCA GGA AC - #C AAG GCT TAT GTG TCT           96                                                                       Ala Glu Lys Asp Tyr Glu Pro Pro Ala Gly Th - #r Lys Ala Tyr Val Ser                        20     - #             25     - #             30                  - - GGA TGG GGA AGA ACA TCG TTC GGT GGC CAA TT - #G TCT AAA AAT CTG CGA          144                                                                       Gly Trp Gly Arg Thr Ser Phe Gly Gly Gln Le - #u Ser Lys Asn Leu Arg                    35         - #         40         - #         45                      - - GGA GTC GAG TTG GAA ATA ATA GAT CTA TTC GA - #T TGT TTC CTT TCC TAC          192                                                                       Gly Val Glu Leu Glu Ile Ile Asp Leu Phe As - #p Cys Phe Leu Ser Tyr                50             - #     55             - #     60                          - - ATG GAT AAA GTA AAC GTG TCC GAA AGG CAA GT - #T TGC GCT GGA ATC CCC          240                                                                       Met Asp Lys Val Asn Val Ser Glu Arg Gln Va - #l Cys Ala Gly Ile Pro            65                 - # 70                 - # 75                 - # 80       - - GTT GTA GGT GGT AAA GAT TCT TGC  CA   - #                  - #                 266                                                                     Val Val Gly Gly Lys Asp Ser Cys                                                                85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:95:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 88 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                              - - Leu Ala Tyr Pro Leu Lys Phe Ser Glu Asp Il - #e Gln Pro Ile Met Met        1               5 - #                 10 - #                 15              - - Ala Glu Lys Asp Tyr Glu Pro Pro Ala Gly Th - #r Lys Ala Tyr Val Ser                   20     - #             25     - #             30                  - - Gly Trp Gly Arg Thr Ser Phe Gly Gly Gln Le - #u Ser Lys Asn Leu Arg               35         - #         40         - #         45                      - - Gly Val Glu Leu Glu Ile Ile Asp Leu Phe As - #p Cys Phe Leu Ser Tyr           50             - #     55             - #     60                          - - Met Asp Lys Val Asn Val Ser Glu Arg Gln Va - #l Cys Ala Gly Ile Pro       65                 - # 70                 - # 75                 - # 80       - - Val Val Gly Gly Lys Asp Ser Cys                                                           85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:96:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 378 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..376                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 3, change G to                        S.  At - # pos. bp 270, change A to W.  At pos. - #aa 1       and aa                                                                                         90, subst - #itute Xaa."                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                              - - GGS GGT ACT TCC CAC CGT GTT GCC CAA ATC GT - #C GTC CAC CCA CAA        TAC       48                                                                    Xaa Gly Thr Ser His Arg Val Ala Gln Ile Va - #l Val His Pro Gln Tyr            1               5 - #                 10 - #                 15              - - AAC GGC AAC ACC AAC ATC AAC GAT GTT GCT GT - #T CTC CGT GTT CAA GAC           96                                                                       Asn Gly Asn Thr Asn Ile Asn Asp Val Ala Va - #l Leu Arg Val Gln Asp                        20     - #             25     - #             30                  - - AAA TTC GTA TTA AAC GGC AGA TCA GTT CGC CC - #C GTT GAC ATG ATC GCT          144                                                                       Lys Phe Val Leu Asn Gly Arg Ser Val Arg Pr - #o Val Asp Met Ile Ala                    35         - #         40         - #         45                      - - TCT GGT GTT GAC ACT CCA GCT GGA GCT CCC CT - #T TAC GTC ACT GGA TGG          192                                                                       Ser Gly Val Asp Thr Pro Ala Gly Ala Pro Le - #u Tyr Val Thr Gly Trp                50             - #     55             - #     60                          - - GGT GCA GTC TAC GAA GGA GGT GCA GGA TCC AC - #C CAA TTA CTA GGA GTA          240                                                                       Gly Ala Val Tyr Glu Gly Gly Ala Gly Ser Th - #r Gln Leu Leu Gly Val            65                 - # 70                 - # 75                 - # 80       - - GGT GTA CCC ATG CGT AGC CAC AAA AAC ACW TG - #T AAC AGC AAA TAC TCC          288                                                                       Gly Val Pro Met Arg Ser His Lys Asn Xaa Cy - #s Asn Ser Lys Tyr Ser                            85 - #                 90 - #                 95              - - CAA TTT GGC GGT GTT GCT CCT AGC ATG ATC TG - #C GCT GGA TTT GAC CAA          336                                                                       Gln Phe Gly Gly Val Ala Pro Ser Met Ile Cy - #s Ala Gly Phe Asp Gln                       100      - #           105      - #           110                  - - GGC GGT AAG GAC GCT TGT CAA GGA GAC TCT GG - #T GGT CCT  T TA               - # 378                                                                    Gly Gly Lys Asp Ala Cys Gln Gly Asp Ser Gl - #y Gly Pro                               115          - #       120          - #       125                      - -  - - (2) INFORMATION FOR SEQ ID NO:97:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 125 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                              - - Xaa Gly Thr Ser His Arg Val Ala Gln Ile Va - #l Val His Pro Gln Tyr        1               5 - #                 10 - #                 15              - - Asn Gly Asn Thr Asn Ile Asn Asp Val Ala Va - #l Leu Arg Val Gln Asp                   20     - #             25     - #             30                  - - Lys Phe Val Leu Asn Gly Arg Ser Val Arg Pr - #o Val Asp Met Ile Ala               35         - #         40         - #         45                      - - Ser Gly Val Asp Thr Pro Ala Gly Ala Pro Le - #u Tyr Val Thr Gly Trp           50             - #     55             - #     60                          - - Gly Ala Val Tyr Glu Gly Gly Ala Gly Ser Th - #r Gln Leu Leu Gly Val       65                 - # 70                 - # 75                 - # 80       - - Gly Val Pro Met Arg Ser His Lys Asn Xaa Cy - #s Asn Ser Lys Tyr Ser                       85 - #                 90 - #                 95              - - Gln Phe Gly Gly Val Ala Pro Ser Met Ile Cy - #s Ala Gly Phe Asp Gln                  100      - #           105      - #           110                  - - Gly Gly Lys Asp Ala Cys Gln Gly Asp Ser Gl - #y Gly Pro                          115          - #       120          - #       125                      - -  - - (2) INFORMATION FOR SEQ ID NO:98:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 252 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..252                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                              - - CGC GCG CCC TGT TGC CCT ACC AGC AGA GAA CG - #A GGA AAC CGA AAC AGG           48                                                                       Arg Ala Pro Cys Cys Pro Thr Ser Arg Glu Ar - #g Gly Asn Arg Asn Arg             1               5 - #                 10 - #                 15              - - GTC ACA CTC ACG GTG ACG GGT TGG GGA ACT AC - #A GAG AGT ACT GAA TCA           96                                                                       Val Thr Leu Thr Val Thr Gly Trp Gly Thr Th - #r Glu Ser Thr Glu Ser                        20     - #             25     - #             30                  - - TCA CAC CAC CTG AAA GAA GTT GAA GTG AAC GC - #T GTA TCT AAT AGT GAA          144                                                                       Ser His His Leu Lys Glu Val Glu Val Asn Al - #a Val Ser Asn Ser Glu                    35         - #         40         - #         45                      - - TGT CAA AGG CCT AAT GAA GAT CTT GCT ACT AT - #A TCA TCA CAT GAG ATA          192                                                                       Cys Gln Arg Pro Asn Glu Asp Leu Ala Thr Il - #e Ser Ser His Glu Ile                50             - #     55             - #     60                          - - TGT GCA AGC GTT CCT GGT GGC GGC AAA GAT TC - #T TGT CAA GGA GAC TCT          240                                                                       Cys Ala Ser Val Pro Gly Gly Gly Lys Asp Se - #r Cys Gln Gly Asp Ser            65                 - # 70                 - # 75                 - # 80       - - GGT GGT CCT TTA            - #                  - #                      - #      252                                                                  Gly Gly Pro Leu                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:99:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 84 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                              - - Arg Ala Pro Cys Cys Pro Thr Ser Arg Glu Ar - #g Gly Asn Arg Asn Arg        1               5 - #                 10 - #                 15              - - Val Thr Leu Thr Val Thr Gly Trp Gly Thr Th - #r Glu Ser Thr Glu Ser                   20     - #             25     - #             30                  - - Ser His His Leu Lys Glu Val Glu Val Asn Al - #a Val Ser Asn Ser Glu               35         - #         40         - #         45                      - - Cys Gln Arg Pro Asn Glu Asp Leu Ala Thr Il - #e Ser Ser His Glu Ile           50             - #     55             - #     60                          - - Cys Ala Ser Val Pro Gly Gly Gly Lys Asp Se - #r Cys Gln Gly Asp Ser       65                 - # 70                 - # 75                 - # 80       - - Gly Gly Pro Leu                                                           - -  - - (2) INFORMATION FOR SEQ ID NO:100:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 225 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..208                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                             - - CCA ATC CAC GAT AGC CAA TAT GCA CTT TTG CA - #G ATA TGG GTC AAG GGT           48                                                                       Pro Ile His Asp Ser Gln Tyr Ala Leu Leu Gl - #n Ile Trp Val Lys Gly             1               5 - #                 10 - #                 15              - - GCA TGT AAG GGT GAT TCC GGT GGC CCC TTA GT - #C ATC AAT GGA CAA CTT           96                                                                       Ala Cys Lys Gly Asp Ser Gly Gly Pro Leu Va - #l Ile Asn Gly Gln Leu                        20     - #             25     - #             30                  - - CAT GGA ATT GTT TCC TGG GGC ATT CCT TGC GC - #T GTC GCA AGC CTG ATG          144                                                                       His Gly Ile Val Ser Trp Gly Ile Pro Cys Al - #a Val Ala Ser Leu Met                    35         - #         40         - #         45                      - - TAT TCA CAA GAG TTT CTC ATT ATG TCG ATT GG - #A TTA AAT CCA AAA TTG          192                                                                       Tyr Ser Gln Glu Phe Leu Ile Met Ser Ile Gl - #y Leu Asn Pro Lys Leu                50             - #     55             - #     60                          - - CCA AAT AAA ATT GTT  T AGAGTATTAA AAAAAAA  - #                  - #            225                                                                     Pro Asn Lys Ile Val                                                            65                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:101:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 69 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                             - - Pro Ile His Asp Ser Gln Tyr Ala Leu Leu Gl - #n Ile Trp Val Lys Gly        1               5 - #                 10 - #                 15              - - Ala Cys Lys Gly Asp Ser Gly Gly Pro Leu Va - #l Ile Asn Gly Gln Leu                   20     - #             25     - #             30                  - - His Gly Ile Val Ser Trp Gly Ile Pro Cys Al - #a Val Ala Ser Leu Met               35         - #         40         - #         45                      - - Tyr Ser Gln Glu Phe Leu Ile Met Ser Ile Gl - #y Leu Asn Pro Lys Leu           50             - #     55             - #     60                          - - Pro Asn Lys Ile Val                                                       65                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:102:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 850 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..758                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                             - - C CTG GTG AAA CTT TTC TTT GTA ATG TAC TGT - #GCT TGT GCA TTA GCA             46                                                                          Leu Val Lys Leu Phe Phe Val Met Tyr C - #ys Ala Cys Ala Leu Ala                 1              - # 5                 - # 10                 - # 15         - - TCG GCA CTG AAG TAC TCC ATC GAT CAT GGT CC - #T CGT ATC ATC GGA GGT           94                                                                       Ser Ala Leu Lys Tyr Ser Ile Asp His Gly Pr - #o Arg Ile Ile Gly Gly                            20 - #                 25 - #                 30              - - GAA GTT GCA GGT GAA GGA TCA GCA CCT TAC CA - #G GTG TCC TTA AGA ACC          142                                                                       Glu Val Ala Gly Glu Gly Ser Ala Pro Tyr Gl - #n Val Ser Leu Arg Thr                        35     - #             40     - #             45                  - - AAG GAA GGA AAT CAT TTT TGC GGT GGA TCA AT - #A CTA AAT AAG CGA TGG          190                                                                       Lys Glu Gly Asn His Phe Cys Gly Gly Ser Il - #e Leu Asn Lys Arg Trp                    50         - #         55         - #         60                      - - GTT GTA ACT GCA GCA CAT TGT CTT GAA CCG GA - #A ATA TTA GAT TCG GTA          238                                                                       Val Val Thr Ala Ala His Cys Leu Glu Pro Gl - #u Ile Leu Asp Ser Val                65             - #     70             - #     75                          - - TAC GTC GGA TCC AAT CAC TTA GAC CGA AAA GG - #C AGA TAT TAC GAC GTA          286                                                                       Tyr Val Gly Ser Asn His Leu Asp Arg Lys Gl - #y Arg Tyr Tyr Asp Val            80                 - # 85                 - # 90                 - # 95       - - GAA CGG TAT ATA ATT CAT GAA AAA TAT ATA GG - #A GAA CTA AAT AAT TTT          334                                                                       Glu Arg Tyr Ile Ile His Glu Lys Tyr Ile Gl - #y Glu Leu Asn Asn Phe                           100  - #               105  - #               110              - - TAT GCT GAC ATC GGT CTA ATA AAA CTT GAT GA - #A GAC TTA GAA TTC AAT          382                                                                       Tyr Ala Asp Ile Gly Leu Ile Lys Leu Asp Gl - #u Asp Leu Glu Phe Asn                       115      - #           120      - #           125                  - - GAC AAA GTC AAG CCA ATA AAA ATT CAT GAA AA - #C ACA ATT CAA GGT GGT          430                                                                       Asp Lys Val Lys Pro Ile Lys Ile His Glu As - #n Thr Ile Gln Gly Gly                   130          - #       135          - #       140                      - - GAA GGG CTT AGA GCA ACA GGT TGG GGA CGT CT - #T GGT GCT GGT CGC CCA          478                                                                       Glu Gly Leu Arg Ala Thr Gly Trp Gly Arg Le - #u Gly Ala Gly Arg Pro               145              - #   150              - #   155                          - - ATT CCT AAT AAA TTG CAG GAG CTA CAA ACA TT - #T GCT TTA AGT GAT AAA          526                                                                       Ile Pro Asn Lys Leu Gln Glu Leu Gln Thr Ph - #e Ala Leu Ser Asp Lys           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - GAT TGT ACA GTA AAA ACT GGT CTT GTA CCA AA - #G TCA CAA CTT TGT        GTT      574                                                                    Asp Cys Thr Val Lys Thr Gly Leu Val Pro Ly - #s Ser Gln Leu Cys Val                          180  - #               185  - #               190              - - TTC CGT GCA TCG GAA AAA GGA GTT TGC TTT GG - #T GAT TCG GGA GGT CCT          622                                                                       Phe Arg Ala Ser Glu Lys Gly Val Cys Phe Gl - #y Asp Ser Gly Gly Pro                       195      - #           200      - #           205                  - - TTG GCA ATC AAT GGT GAA CTT GTT GGT GTT AC - #T TCA TTC ATT ATG GGA          670                                                                       Leu Ala Ile Asn Gly Glu Leu Val Gly Val Th - #r Ser Phe Ile Met Gly                   210          - #       215          - #       220                      - - ACA TGT GGA GGA GGA CAT CCT GAT GTC TTC GG - #T CGA GTC CTT GAC TTC          718                                                                       Thr Cys Gly Gly Gly His Pro Asp Val Phe Gl - #y Arg Val Leu Asp Phe               225              - #   230              - #   235                          - - AAA CCA TGG ATT GAT TCT CAT ATG GCA AAT GA - #C GGC GCA T AATTCTTTTA         768                                                                       Lys Pro Trp Ile Asp Ser His Met Ala Asn As - #p Gly Ala                       240                 2 - #45                 2 - #50                            - - TTTAATAATG ATTGAATGTA AAATTATAAA CAAATTGTAA ATTGCATAAA TG -             #ATATAAAT    828                                                                 - - GCAGGAAATT CGAAAAAAAA AA           - #                  - #                    850                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:103:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 252 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                             - - Leu Val Lys Leu Phe Phe Val Met Tyr Cys Al - #a Cys Ala Leu Ala Ser        1               5 - #                 10 - #                 15              - - Ala Leu Lys Tyr Ser Ile Asp His Gly Pro Ar - #g Ile Ile Gly Gly Glu                   20     - #             25     - #             30                  - - Val Ala Gly Glu Gly Ser Ala Pro Tyr Gln Va - #l Ser Leu Arg Thr Lys               35         - #         40         - #         45                      - - Glu Gly Asn His Phe Cys Gly Gly Ser Ile Le - #u Asn Lys Arg Trp Val           50             - #     55             - #     60                          - - Val Thr Ala Ala His Cys Leu Glu Pro Glu Il - #e Leu Asp Ser Val Tyr       65                 - # 70                 - # 75                 - # 80       - - Val Gly Ser Asn His Leu Asp Arg Lys Gly Ar - #g Tyr Tyr Asp Val Glu                       85 - #                 90 - #                 95              - - Arg Tyr Ile Ile His Glu Lys Tyr Ile Gly Gl - #u Leu Asn Asn Phe Tyr                  100      - #           105      - #           110                  - - Ala Asp Ile Gly Leu Ile Lys Leu Asp Glu As - #p Leu Glu Phe Asn Asp              115          - #       120          - #       125                      - - Lys Val Lys Pro Ile Lys Ile His Glu Asn Th - #r Ile Gln Gly Gly Glu          130              - #   135              - #   140                          - - Gly Leu Arg Ala Thr Gly Trp Gly Arg Leu Gl - #y Ala Gly Arg Pro Ile      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Pro Asn Lys Leu Gln Glu Leu Gln Thr Phe Al - #a Leu Ser Asp Lys        Asp                                                                                             165  - #               170  - #               175             - - Cys Thr Val Lys Thr Gly Leu Val Pro Lys Se - #r Gln Leu Cys Val Phe                  180      - #           185      - #           190                  - - Arg Ala Ser Glu Lys Gly Val Cys Phe Gly As - #p Ser Gly Gly Pro Leu              195          - #       200          - #       205                      - - Ala Ile Asn Gly Glu Leu Val Gly Val Thr Se - #r Phe Ile Met Gly Thr          210              - #   215              - #   220                          - - Cys Gly Gly Gly His Pro Asp Val Phe Gly Ar - #g Val Leu Asp Phe Lys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Pro Trp Ile Asp Ser His Met Ala Asn Asp Gl - #y Ala                                      245  - #               250                                     - -  - - (2) INFORMATION FOR SEQ ID NO:104:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 252 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..251                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 4, change A to                       R.  At - # pos. aa 2, substitute Xaa."                          - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                             - - GCA RGG ATT CGG CAC GAG AAT TTA TTA AGC GC - #A TTA TTT GCA AGT GTA           48                                                                       Ala Xaa Ile Arg His Glu Asn Leu Leu Ser Al - #a Leu Phe Ala Ser Val             1               5 - #                 10 - #                 15              - - ATT TGC TCC TTT AAC GCG GAA GTA CAA AAT CG - #A ATC GTT GGT GGC AAT           96                                                                       Ile Cys Ser Phe Asn Ala Glu Val Gln Asn Ar - #g Ile Val Gly Gly Asn                        20     - #             25     - #             30                  - - GAT GTA AGT ATT TCA AAA ATT GGG TGG CAA GT - #A TCT ATT CAA AGT AAT          144                                                                       Asp Val Ser Ile Ser Lys Ile Gly Trp Gln Va - #l Ser Ile Gln Ser Asn                    35         - #         40         - #         45                      - - AAC CAA CAT TTC TGT GGT GGT TCA ATC ATT GC - #T AAA GAT TGG GTA CTG          192                                                                       Asn Gln His Phe Cys Gly Gly Ser Ile Ile Al - #a Lys Asp Trp Val Leu                50             - #     55             - #     60                          - - ACT TCT TCT CAA TGC GTC GTG GAC AAA CAA AG - #T CCA CCG AAG GAT TTA          240                                                                       Thr Ser Ser Gln Cys Val Val Asp Lys Gln Se - #r Pro Pro Lys Asp Leu            65                 - # 70                 - # 75                 - # 80       - - ACT GTT CGT  GT T          - #                  - #                      - #      252                                                                  Thr Val Arg                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:105:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 83 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                             - - Ala Xaa Ile Arg His Glu Asn Leu Leu Ser Al - #a Leu Phe Ala Ser Val        1               5 - #                 10 - #                 15              - - Ile Cys Ser Phe Asn Ala Glu Val Gln Asn Ar - #g Ile Val Gly Gly Asn                   20     - #             25     - #             30                  - - Asp Val Ser Ile Ser Lys Ile Gly Trp Gln Va - #l Ser Ile Gln Ser Asn               35         - #         40         - #         45                      - - Asn Gln His Phe Cys Gly Gly Ser Ile Ile Al - #a Lys Asp Trp Val Leu           50             - #     55             - #     60                          - - Thr Ser Ser Gln Cys Val Val Asp Lys Gln Se - #r Pro Pro Lys Asp Leu       65                 - # 70                 - # 75                 - # 80       - - Thr Val Arg                                                               - -  - - (2) INFORMATION FOR SEQ ID NO:106:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 534 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..534                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:                             - - TCA AAT CGG ATT GTT AAT GGA GTT AAT GCC AA - #A AAC GGT TCT GCT CCA           48                                                                       Ser Asn Arg Ile Val Asn Gly Val Asn Ala Ly - #s Asn Gly Ser Ala Pro             1               5 - #                 10 - #                 15              - - TAT ATG GCT TCT CTA AGA GAT GTT ATG GAA AC - #C ATT TCT GTG GAG CAT           96                                                                       Tyr Met Ala Ser Leu Arg Asp Val Met Glu Th - #r Ile Ser Val Glu His                        20     - #             25     - #             30                  - - CGA TAT TGG ATG AAC CGC TGG ATT CTT ACT GC - #T GCC CAT TGC CTT ACT          144                                                                       Arg Tyr Trp Met Asn Arg Trp Ile Leu Thr Al - #a Ala His Cys Leu Thr                    35         - #         40         - #         45                      - - GAC GGT TAT CTA GAT ACA GTC TAC GTT GGT TC - #A AAT CAT CTT TCT GGC          192                                                                       Asp Gly Tyr Leu Asp Thr Val Tyr Val Gly Se - #r Asn His Leu Ser Gly                50             - #     55             - #     60                          - - GAC GGA GAG TAC TAC AAT GTA GAA GAA CAA GT - #C ATC CAT GAT AAA TAT          240                                                                       Asp Gly Glu Tyr Tyr Asn Val Glu Glu Gln Va - #l Ile His Asp Lys Tyr            65                 - # 70                 - # 75                 - # 80       - - TTT GGT CAA ACA ACC GGC TTC AAA AAT GAT AT - #T GCT CTC GTC AAA GTT          288                                                                       Phe Gly Gln Thr Thr Gly Phe Lys Asn Asp Il - #e Ala Leu Val Lys Val                            85 - #                 90 - #                 95              - - TCT AGT GCT ATA AAA CTT AGC AAA AAT GTT CG - #T CCC ATC AAA TTG CAC          336                                                                       Ser Ser Ala Ile Lys Leu Ser Lys Asn Val Ar - #g Pro Ile Lys Leu His                       100      - #           105      - #           110                  - - AAA GAT TTT ATA CGC GGA GGT GAA AAA TTG AA - #A ATT ACT GGA TGG GGA          384                                                                       Lys Asp Phe Ile Arg Gly Gly Glu Lys Leu Ly - #s Ile Thr Gly Trp Gly                   115          - #       120          - #       125                      - - TTG ACC AAT CAA ACT CAT GGT GAA GTT CCT GA - #T GCT CTT CAA GAG TTA          432                                                                       Leu Thr Asn Gln Thr His Gly Glu Val Pro As - #p Ala Leu Gln Glu Leu               130              - #   135              - #   140                          - - CAG GTA GAA GCA CTT TCT AAC TCT AAA TGC AA - #G GCA ATT ACT GGT GTC          480                                                                       Gln Val Glu Ala Leu Ser Asn Ser Lys Cys Ly - #s Ala Ile Thr Gly Val           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - CAT CTT CCT GCT CAT CTC TGC ACC TTC AGA GC - #T CCT CAA AAG GGT        GTA      528                                                                    His Leu Pro Ala His Leu Cys Thr Phe Arg Al - #a Pro Gln Lys Gly Val                          165  - #               170  - #               175              - - TGC CAG                - #                  - #                  -      #          534                                                                  Cys Gln                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:107:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 178 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:                             - - Ser Asn Arg Ile Val Asn Gly Val Asn Ala Ly - #s Asn Gly Ser Ala        Pro                                                                               1               5 - #                 10 - #                 15             - - Tyr Met Ala Ser Leu Arg Asp Val Met Glu Th - #r Ile Ser Val Glu His                   20     - #             25     - #             30                  - - Arg Tyr Trp Met Asn Arg Trp Ile Leu Thr Al - #a Ala His Cys Leu Thr               35         - #         40         - #         45                      - - Asp Gly Tyr Leu Asp Thr Val Tyr Val Gly Se - #r Asn His Leu Ser Gly           50             - #     55             - #     60                          - - Asp Gly Glu Tyr Tyr Asn Val Glu Glu Gln Va - #l Ile His Asp Lys Tyr       65                 - # 70                 - # 75                 - # 80       - - Phe Gly Gln Thr Thr Gly Phe Lys Asn Asp Il - #e Ala Leu Val Lys Val                       85 - #                 90 - #                 95              - - Ser Ser Ala Ile Lys Leu Ser Lys Asn Val Ar - #g Pro Ile Lys Leu His                  100      - #           105      - #           110                  - - Lys Asp Phe Ile Arg Gly Gly Glu Lys Leu Ly - #s Ile Thr Gly Trp Gly              115          - #       120          - #       125                      - - Leu Thr Asn Gln Thr His Gly Glu Val Pro As - #p Ala Leu Gln Glu Leu          130              - #   135              - #   140                          - - Gln Val Glu Ala Leu Ser Asn Ser Lys Cys Ly - #s Ala Ile Thr Gly Val      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - His Leu Pro Ala His Leu Cys Thr Phe Arg Al - #a Pro Gln Lys Gly        Val                                                                                             165  - #               170  - #               175             - - Cys Gln                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:108:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 359 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..359                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108:                             - - GGG TTC GAA TTT GTG GAT CGA AAA GGC AGA TA - #T TAC GAT GTA GAA AGA           48                                                                       Gly Phe Glu Phe Val Asp Arg Lys Gly Arg Ty - #r Tyr Asp Val Glu Arg             1               5 - #                 10 - #                 15              - - TTT GTG ATG CAC CAT AAT TAT ACT GGA AAG AT - #A GTT GCC AAT GTC GCT           96                                                                       Phe Val Met His His Asn Tyr Thr Gly Lys Il - #e Val Ala Asn Val Ala                        20     - #             25     - #             30                  - - GAT ATA GGT CTA ATA AAA CTA GCA GAA GAT AT - #A AAA TTC AGT GAC AAG          144                                                                       Asp Ile Gly Leu Ile Lys Leu Ala Glu Asp Il - #e Lys Phe Ser Asp Lys                    35         - #         40         - #         45                      - - GTA CAA CCT GTA AAA ATT CAT CAA ACT CAA AT - #C AAG GGC GGA GAG ATT          192                                                                       Val Gln Pro Val Lys Ile His Gln Thr Gln Il - #e Lys Gly Gly Glu Ile                50             - #     55             - #     60                          - - TGC AAA GCT ACT GGA TGG GGC AGG TTG GGT GC - #T GAT CAG CCT GTA CCA          240                                                                       Cys Lys Ala Thr Gly Trp Gly Arg Leu Gly Al - #a Asp Gln Pro Val Pro            65                 - # 70                 - # 75                 - # 80       - - AAT AAA TTA CAA CAA TTG GAG ACA ATT GCT AT - #T AGT GAT GAG AAA TGT          288                                                                       Asn Lys Leu Gln Gln Leu Glu Thr Ile Ala Il - #e Ser Asp Glu Lys Cys                            85 - #                 90 - #                 95              - - TAT GCA GAT ACA GGG TTT TTA GAA CCT ACA TC - #T CAA ATA TGT GTA TTC          336                                                                       Tyr Ala Asp Thr Gly Phe Leu Glu Pro Thr Se - #r Gln Ile Cys Val Phe                       100      - #           105      - #           110                  - - AGT GCA TTT GGA AAA GGA GTT  GT     - #                  - #                   359                                                                     Ser Ala Phe Gly Lys Gly Val                                                           115                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:109:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 119 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109:                             - - Gly Phe Glu Phe Val Asp Arg Lys Gly Arg Ty - #r Tyr Asp Val Glu Arg        1               5 - #                 10 - #                 15              - - Phe Val Met His His Asn Tyr Thr Gly Lys Il - #e Val Ala Asn Val Ala                   20     - #             25     - #             30                  - - Asp Ile Gly Leu Ile Lys Leu Ala Glu Asp Il - #e Lys Phe Ser Asp Lys               35         - #         40         - #         45                      - - Val Gln Pro Val Lys Ile His Gln Thr Gln Il - #e Lys Gly Gly Glu Ile           50             - #     55             - #     60                          - - Cys Lys Ala Thr Gly Trp Gly Arg Leu Gly Al - #a Asp Gln Pro Val Pro       65                 - # 70                 - # 75                 - # 80       - - Asn Lys Leu Gln Gln Leu Glu Thr Ile Ala Il - #e Ser Asp Glu Lys Cys                       85 - #                 90 - #                 95              - - Tyr Ala Asp Thr Gly Phe Leu Glu Pro Thr Se - #r Gln Ile Cys Val Phe                  100      - #           105      - #           110                  - - Ser Ala Phe Gly Lys Gly Val                                                      115                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:110:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 841 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..746                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 306, change A to                      M.  At - # pos. bp 382, change A to M.  At pos. - #bp.        394,                                                                                           change G - #to S.  At pos. aa 102, 127 and 131, subst -      #itute                                                                                         Xaa."                                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:                             - - C TTA GCA ATT GTA TGT GCT CTC GCT GTC TGC - #ACA TTC GGT GCC AGT             46                                                                         Leu Ala Ile Val Cys Ala Leu Ala Val C - #ys Thr Phe Gly Ala Ser                 1              - # 5                 - # 10                 - # 15         - - GTT CCA GAA TCA TGG AAA AGA TTA GAT AGT AG - #A ATC GTA GGA GGA CAC           94                                                                       Val Pro Glu Ser Trp Lys Arg Leu Asp Ser Ar - #g Ile Val Gly Gly His                            20 - #                 25 - #                 30              - - GAT ACC AGC ATC GAT AAA CAC CCT CAT CAA GT - #A TCT TTA TTG TAC TCC          142                                                                       Asp Thr Ser Ile Asp Lys His Pro His Gln Va - #l Ser Leu Leu Tyr Ser                        35     - #             40     - #             45                  - - AGC CAC AAT TGT GGT GGT TCC TTG ATT GCC AA - #A AAC TGG TGG GTT TTG          190                                                                       Ser His Asn Cys Gly Gly Ser Leu Ile Ala Ly - #s Asn Trp Trp Val Leu                    50         - #         55         - #         60                      - - ACT GCA GCT CAT TGC ATT GGA GTT AAC AAA TA - #C AAT GTC CGT GTA GGA          238                                                                       Thr Ala Ala His Cys Ile Gly Val Asn Lys Ty - #r Asn Val Arg Val Gly                65             - #     70             - #     75                          - - AGT TCC ATC GTA AAC AGC GGT GGT ATC TTG CA - #T AAA GTT AAA AAC CAT          286                                                                       Ser Ser Ile Val Asn Ser Gly Gly Ile Leu Hi - #s Lys Val Lys Asn His            80                 - # 85                 - # 90                 - # 95       - - TAC AGA CAT CCA AAA TAC AMC GCA GCT GCT AT - #T GAC TTT GAT TAC GCA          334                                                                       Tyr Arg His Pro Lys Tyr Xaa Ala Ala Ala Il - #e Asp Phe Asp Tyr Ala                           100  - #               105  - #               110              - - CTC TTA GAA CTC GAA ACT CCT GTT CAA CTC AC - #A AAT GAT GTG TCC ATM          382                                                                       Leu Leu Glu Leu Glu Thr Pro Val Gln Leu Th - #r Asn Asp Val Ser Xaa                       115      - #           120      - #           125                  - - ATA AAA TTG GTS GAT GAA GGA GTA GAT CTT AA - #A CCT GGT ACC TTG TTA          430                                                                       Ile Lys Leu Xaa Asp Glu Gly Val Asp Leu Ly - #s Pro Gly Thr Leu Leu                   130          - #       135          - #       140                      - - ACT GTT ACT GGA TGG GGA TCA ACT GGA AAT GG - #A CCT TCA ACC AAT GTT          478                                                                       Thr Val Thr Gly Trp Gly Ser Thr Gly Asn Gl - #y Pro Ser Thr Asn Val               145              - #   150              - #   155                          - - TTG CAA GAA GTT CAA GTA CCA CAT GTC GAC CA - #A ACC ACT TGC TCC AAA          526                                                                       Leu Gln Glu Val Gln Val Pro His Val Asp Gl - #n Thr Thr Cys Ser Lys           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - TCT TAC CCA GGA AGT TTG ACT GAT CGT ATG TT - #C TGC GCT GGT TAT        TTG      574                                                                    Ser Tyr Pro Gly Ser Leu Thr Asp Arg Met Ph - #e Cys Ala Gly Tyr Leu                          180  - #               185  - #               190              - - GGA CAA GGA GGC AAG GAC TCA TGC CAA GGT GA - #T TCT GGT GGC CCA GTT          622                                                                       Gly Gln Gly Gly Lys Asp Ser Cys Gln Gly As - #p Ser Gly Gly Pro Val                       195      - #           200      - #           205                  - - GTT GTC AAT GGT GTT CAA CAT GGA ATT GTC TC - #A TGG GGT CGT GGT TGT          670                                                                       Val Val Asn Gly Val Gln His Gly Ile Val Se - #r Trp Gly Arg Gly Cys                   210          - #       215          - #       220                      - - GCA CTT CCT GAT TAT CCT GGA GTT TAC TCT AA - #A ATC TCT ACC GCT CGC          718                                                                       Ala Leu Pro Asp Tyr Pro Gly Val Tyr Ser Ly - #s Ile Ser Thr Ala Arg               225              - #   230              - #   235                          - - AGC TGG ATC AAG GAA GTG TCT GGT GTT T AATT - #TATTCT TGAAATCTCT              766                                                                       Ser Trp Ile Lys Glu Val Ser Gly Val                                           240                 2 - #45                                                    - - ATTTTGTATT ATTTATGTAT ATAGTAAGAG TTGTAAATAT AAATAGTTAC AT -             #CTAAAAAA    826                                                                 - - AAAAAAAAAA AAAAA              - #                  - #                      - #   841                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:111:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 248 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:                             - - Leu Ala Ile Val Cys Ala Leu Ala Val Cys Th - #r Phe Gly Ala Ser Val        1               5 - #                 10 - #                 15              - - Pro Glu Ser Trp Lys Arg Leu Asp Ser Arg Il - #e Val Gly Gly His Asp                   20     - #             25     - #             30                  - - Thr Ser Ile Asp Lys His Pro His Gln Val Se - #r Leu Leu Tyr Ser Ser               35         - #         40         - #         45                      - - His Asn Cys Gly Gly Ser Leu Ile Ala Lys As - #n Trp Trp Val Leu Thr           50             - #     55             - #     60                          - - Ala Ala His Cys Ile Gly Val Asn Lys Tyr As - #n Val Arg Val Gly Ser       65                 - # 70                 - # 75                 - # 80       - - Ser Ile Val Asn Ser Gly Gly Ile Leu His Ly - #s Val Lys Asn His Tyr                       85 - #                 90 - #                 95              - - Arg His Pro Lys Tyr Xaa Ala Ala Ala Ile As - #p Phe Asp Tyr Ala Leu                  100      - #           105      - #           110                  - - Leu Glu Leu Glu Thr Pro Val Gln Leu Thr As - #n Asp Val Ser Xaa Ile              115          - #       120          - #       125                      - - Lys Leu Xaa Asp Glu Gly Val Asp Leu Lys Pr - #o Gly Thr Leu Leu Thr          130              - #   135              - #   140                          - - Val Thr Gly Trp Gly Ser Thr Gly Asn Gly Pr - #o Ser Thr Asn Val Leu      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gln Glu Val Gln Val Pro His Val Asp Gln Th - #r Thr Cys Ser Lys        Ser                                                                                             165  - #               170  - #               175             - - Tyr Pro Gly Ser Leu Thr Asp Arg Met Phe Cy - #s Ala Gly Tyr Leu Gly                  180      - #           185      - #           190                  - - Gln Gly Gly Lys Asp Ser Cys Gln Gly Asp Se - #r Gly Gly Pro Val Val              195          - #       200          - #       205                      - - Val Asn Gly Val Gln His Gly Ile Val Ser Tr - #p Gly Arg Gly Cys Ala          210              - #   215              - #   220                          - - Leu Pro Asp Tyr Pro Gly Val Tyr Ser Lys Il - #e Ser Thr Ala Arg Ser      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Trp Ile Lys Glu Val Ser Gly Val                                                          245                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:112:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1580 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..1491                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:                             - - GG CAC GAG TTT TGT GCG AGT GTC AGA TAT TGC - # AGC TCT ATG AGT AAC            47                                                                         His Glu Phe Cys Ala Ser Val Arg Tyr - #Cys Ser Ser Met Ser Asn                  1             - #  5                - #  10                - #  15        - - AAG AAA GGA TTA GTA CTG GGC ATC TAC GAC AA - #T GAA TTC GAT AAA AAA           95                                                                       Lys Lys Gly Leu Val Leu Gly Ile Tyr Asp As - #n Glu Phe Asp Lys Lys                            20 - #                 25 - #                 30              - - ATA AGG TTA ACG CCA ACT GCT GAA CAA TTC AA - #T CGG CGA TTG CAG GGG          143                                                                       Ile Arg Leu Thr Pro Thr Ala Glu Gln Phe As - #n Arg Arg Leu Gln Gly                        35     - #             40     - #             45                  - - CGT TTA CTA GAT CTA ATT CAT TTG AGT GGA CC - #C ATT AAA TTG GGC AAG          191                                                                       Arg Leu Leu Asp Leu Ile His Leu Ser Gly Pr - #o Ile Lys Leu Gly Lys                    50         - #         55         - #         60                      - - AGC CGT ATT TTC TGG GAT CTC GAT GAA TTC GG - #C GCA GTT GCA GTT GCA          239                                                                       Ser Arg Ile Phe Trp Asp Leu Asp Glu Phe Gl - #y Ala Val Ala Val Ala                65             - #     70             - #     75                          - - GGT TTG GGA AAT CAC TCC CCC TGC GAA CTC CT - #G GAA GAA CTC GAT GTT          287                                                                       Gly Leu Gly Asn His Ser Pro Cys Glu Leu Le - #u Glu Glu Leu Asp Val            80                 - # 85                 - # 90                 - # 95       - - TTG CGC GAA AAT GCC AGA ATA GCT GCC GGT GC - #T GGT TGC CAA GCT CTT          335                                                                       Leu Arg Glu Asn Ala Arg Ile Ala Ala Gly Al - #a Gly Cys Gln Ala Leu                           100  - #               105  - #               110              - - GCC GCC GAT GGA ATC ACT ACC ATT AGC GTT GA - #A GTA TGG AGC ACC CGG          383                                                                       Ala Ala Asp Gly Ile Thr Thr Ile Ser Val Gl - #u Val Trp Ser Thr Arg                       115      - #           120      - #           125                  - - AGG CGG CCA TGC GAA GGT GCA ATA CTA TCG AC - #G TTC AAA TTC AGG TCA          431                                                                       Arg Arg Pro Cys Glu Gly Ala Ile Leu Ser Th - #r Phe Lys Phe Arg Ser                   130          - #       135          - #       140                      - - ACA GAA GAA GAG TCC AAG TGT AAG CCG ATA CC - #T ACC ATA ACC CCT TAC          479                                                                       Thr Glu Glu Glu Ser Lys Cys Lys Pro Ile Pr - #o Thr Ile Thr Pro Tyr               145              - #   150              - #   155                          - - TGC CTT CAA GAT AAA GAT GCT CCA TTA TGG GA - #A CTT GGC CAA GTA TCA          527                                                                       Cys Leu Gln Asp Lys Asp Ala Pro Leu Trp Gl - #u Leu Gly Gln Val Ser           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - GCA GCA GCT CAA AAC TGG GCT CGT ACA TTG AT - #G GAT ACA CCA GCA        AAT      575                                                                    Ala Ala Ala Gln Asn Trp Ala Arg Thr Leu Me - #t Asp Thr Pro Ala Asn                          180  - #               185  - #               190              - - CAA ATG ACA CCA TTT TTG TTC GCC GAA GCC GC - #C AAA GAA AAT TTA GTG          623                                                                       Gln Met Thr Pro Phe Leu Phe Ala Glu Ala Al - #a Lys Glu Asn Leu Val                       195      - #           200      - #           205                  - - CCA TTA GGA GTG AAA GTT GAA GCT AGA GAT CG - #G AAA TGG GCG GTA AGC          671                                                                       Pro Leu Gly Val Lys Val Glu Ala Arg Asp Ar - #g Lys Trp Ala Val Ser                   210          - #       215          - #       220                      - - ATG AAA ATG GGA TCC TTC TTG TCT GTC GCT CG - #T GGC TCC AAT GAA CCA          719                                                                       Met Lys Met Gly Ser Phe Leu Ser Val Ala Ar - #g Gly Ser Asn Glu Pro               225              - #   230              - #   235                          - - CCA GTT TTT CTT GAA ATT TCT TAT TGT GGT GG - #T CCA AAA GAT GAG GCA          767                                                                       Pro Val Phe Leu Glu Ile Ser Tyr Cys Gly Gl - #y Pro Lys Asp Glu Ala           240                 2 - #45                 2 - #50                 2 -      #55                                                                              - - CCG TTC GCA CTT GTT GGA AAG GGT GTC ACT TT - #C GAT ACT GGC GGT        ATT      815                                                                    Pro Phe Ala Leu Val Gly Lys Gly Val Thr Ph - #e Asp Thr Gly Gly Ile                          260  - #               265  - #               270              - - AGC ATC AAA CCG AGT GCA TCC ATG GAC GAA AT - #G CGT GGA GAT ATG GGA          863                                                                       Ser Ile Lys Pro Ser Ala Ser Met Asp Glu Me - #t Arg Gly Asp Met Gly                       275      - #           280      - #           285                  - - GGA GCT GCT TGC GTT GTT TCT ACA TTG GCA CA - #A TTG AAA GCA CCA GTC          911                                                                       Gly Ala Ala Cys Val Val Ser Thr Leu Ala Gl - #n Leu Lys Ala Pro Val                   290          - #       295          - #       300                      - - AAC GTC GTC GGT CTT ATC CCC TTA ACC GAG AA - #T ATG CCA GGT GGT AAA          959                                                                       Asn Val Val Gly Leu Ile Pro Leu Thr Glu As - #n Met Pro Gly Gly Lys               305              - #   310              - #   315                          - - GCA ACA AAA CCT GGT GAC GTC GTT GTT GCG AT - #G AAT GGG AAA TCG ATT         1007                                                                       Ala Thr Lys Pro Gly Asp Val Val Val Ala Me - #t Asn Gly Lys Ser Ile           320                 3 - #25                 3 - #30                 3 -      #35                                                                              - - TGC GTG GAC AAT ACA GAT GCT GAA GGC CGT TT - #G ATT TTA GCT GAC        GCT     1055                                                                    Cys Val Asp Asn Thr Asp Ala Glu Gly Arg Le - #u Ile Leu Ala Asp Ala                          340  - #               345  - #               350              - - TTA TGT TAC TCG GCA CAC TTC AAG CCA AAA TG - #G GTT CTA GAT ATA GCT         1103                                                                       Leu Cys Tyr Ser Ala His Phe Lys Pro Lys Tr - #p Val Leu Asp Ile Ala                       355      - #           360      - #           365                  - - ACA TTG ACT GGA GCT ATG AGA GTT GCT CTA GG - #T GAT TGT GCT ACT GGT         1151                                                                       Thr Leu Thr Gly Ala Met Arg Val Ala Leu Gl - #y Asp Cys Ala Thr Gly                   370          - #       375          - #       380                      - - GTA TTT TCT TCA TGC GAT AAT CTC TGG AAC AC - #A CTG CAC GAA GCT GGT         1199                                                                       Val Phe Ser Ser Cys Asp Asn Leu Trp Asn Th - #r Leu His Glu Ala Gly               385              - #   390              - #   395                          - - AGA GTA ACT GGA GAT AGA ATG TGG AGA TTC CC - #T CTT TTT AAG CAC TAC         1247                                                                       Arg Val Thr Gly Asp Arg Met Trp Arg Phe Pr - #o Leu Phe Lys His Tyr           400                 4 - #05                 4 - #10                 4 -      #15                                                                              - - GCG AAT CGT GTA ACA GAA TAT TCC GGT TAC GA - #T GTG AAC AAC ATA        GGA     1295                                                                    Ala Asn Arg Val Thr Glu Tyr Ser Gly Tyr As - #p Val Asn Asn Ile Gly                          420  - #               425  - #               430              - - AAG GGC AAA GGG GGA GGC AGT TGC CTA GCA GC - #T GCT TTC CTT AAT CAG         1343                                                                       Lys Gly Lys Gly Gly Gly Ser Cys Leu Ala Al - #a Ala Phe Leu Asn Gln                       435      - #           440      - #           445                  - - TTT AGA CCT GAG GAC GTA CCC TGG ATG CAC TT - #G GAC ATT GCT GGA GTA         1391                                                                       Phe Arg Pro Glu Asp Val Pro Trp Met His Le - #u Asp Ile Ala Gly Val                   450          - #       455          - #       460                      - - ATG AGC GAT TGC TCT GAT CAG TCC TAT CTT CC - #T AAG GGA ATG ACA GGG         1439                                                                       Met Ser Asp Cys Ser Asp Gln Ser Tyr Leu Pr - #o Lys Gly Met Thr Gly               465              - #   470              - #   475                          - - CGA CCT ACC AGA ACT CTT GTT CAA TTT ATT CA - #A AGC CAA AAG CGT CAT         1487                                                                       Arg Pro Thr Arg Thr Leu Val Gln Phe Ile Gl - #n Ser Gln Lys Arg His           480                 4 - #85                 4 - #90                 4 -      #95                                                                              - - TCT T GAATGCATTT AAGAAACATA TGTCACCTAA CACCTTTACA TGCG - #CACCTC            1541                                                                      Ser                                                                            - - TGCTTAGCCA AACCACTTTT GTACAAAACC ATATTTTTA      - #                      - #  1580                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:113:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 496 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:                             - - His Glu Phe Cys Ala Ser Val Arg Tyr Cys Se - #r Ser Met Ser Asn Lys        1               5 - #                 10 - #                 15              - - Lys Gly Leu Val Leu Gly Ile Tyr Asp Asn Gl - #u Phe Asp Lys Lys Ile                   20     - #             25     - #             30                  - - Arg Leu Thr Pro Thr Ala Glu Gln Phe Asn Ar - #g Arg Leu Gln Gly Arg               35         - #         40         - #         45                      - - Leu Leu Asp Leu Ile His Leu Ser Gly Pro Il - #e Lys Leu Gly Lys Ser           50             - #     55             - #     60                          - - Arg Ile Phe Trp Asp Leu Asp Glu Phe Gly Al - #a Val Ala Val Ala Gly       65                 - # 70                 - # 75                 - # 80       - - Leu Gly Asn His Ser Pro Cys Glu Leu Leu Gl - #u Glu Leu Asp Val Leu                       85 - #                 90 - #                 95              - - Arg Glu Asn Ala Arg Ile Ala Ala Gly Ala Gl - #y Cys Gln Ala Leu Ala                  100      - #           105      - #           110                  - - Ala Asp Gly Ile Thr Thr Ile Ser Val Glu Va - #l Trp Ser Thr Arg Arg              115          - #       120          - #       125                      - - Arg Pro Cys Glu Gly Ala Ile Leu Ser Thr Ph - #e Lys Phe Arg Ser Thr          130              - #   135              - #   140                          - - Glu Glu Glu Ser Lys Cys Lys Pro Ile Pro Th - #r Ile Thr Pro Tyr Cys      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Leu Gln Asp Lys Asp Ala Pro Leu Trp Glu Le - #u Gly Gln Val Ser        Ala                                                                                             165  - #               170  - #               175             - - Ala Ala Gln Asn Trp Ala Arg Thr Leu Met As - #p Thr Pro Ala Asn Gln                  180      - #           185      - #           190                  - - Met Thr Pro Phe Leu Phe Ala Glu Ala Ala Ly - #s Glu Asn Leu Val Pro              195          - #       200          - #       205                      - - Leu Gly Val Lys Val Glu Ala Arg Asp Arg Ly - #s Trp Ala Val Ser Met          210              - #   215              - #   220                          - - Lys Met Gly Ser Phe Leu Ser Val Ala Arg Gl - #y Ser Asn Glu Pro Pro      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Val Phe Leu Glu Ile Ser Tyr Cys Gly Gly Pr - #o Lys Asp Glu Ala        Pro                                                                                             245  - #               250  - #               255             - - Phe Ala Leu Val Gly Lys Gly Val Thr Phe As - #p Thr Gly Gly Ile Ser                  260      - #           265      - #           270                  - - Ile Lys Pro Ser Ala Ser Met Asp Glu Met Ar - #g Gly Asp Met Gly Gly              275          - #       280          - #       285                      - - Ala Ala Cys Val Val Ser Thr Leu Ala Gln Le - #u Lys Ala Pro Val Asn          290              - #   295              - #   300                          - - Val Val Gly Leu Ile Pro Leu Thr Glu Asn Me - #t Pro Gly Gly Lys Ala      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Thr Lys Pro Gly Asp Val Val Val Ala Met As - #n Gly Lys Ser Ile        Cys                                                                                             325  - #               330  - #               335             - - Val Asp Asn Thr Asp Ala Glu Gly Arg Leu Il - #e Leu Ala Asp Ala Leu                  340      - #           345      - #           350                  - - Cys Tyr Ser Ala His Phe Lys Pro Lys Trp Va - #l Leu Asp Ile Ala Thr              355          - #       360          - #       365                      - - Leu Thr Gly Ala Met Arg Val Ala Leu Gly As - #p Cys Ala Thr Gly Val          370              - #   375              - #   380                          - - Phe Ser Ser Cys Asp Asn Leu Trp Asn Thr Le - #u His Glu Ala Gly Arg      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Val Thr Gly Asp Arg Met Trp Arg Phe Pro Le - #u Phe Lys His Tyr        Ala                                                                                             405  - #               410  - #               415             - - Asn Arg Val Thr Glu Tyr Ser Gly Tyr Asp Va - #l Asn Asn Ile Gly Lys                  420      - #           425      - #           430                  - - Gly Lys Gly Gly Gly Ser Cys Leu Ala Ala Al - #a Phe Leu Asn Gln Phe              435          - #       440          - #       445                      - - Arg Pro Glu Asp Val Pro Trp Met His Leu As - #p Ile Ala Gly Val Met          450              - #   455              - #   460                          - - Ser Asp Cys Ser Asp Gln Ser Tyr Leu Pro Ly - #s Gly Met Thr Gly Arg      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Pro Thr Arg Thr Leu Val Gln Phe Ile Gln Se - #r Gln Lys Arg His        Ser                                                                                             485  - #               490  - #               495             - -  - - (2) INFORMATION FOR SEQ ID NO:114:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 32 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:                             - - GAGCTCTCGA GAGTTGTTGG AGGACTGGAA GC       - #                  - #              32                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:115:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115:                             - - GGACCTCGAG AATTAGTTAT TTTCCATGGT C        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:116:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:                             - - GAGCTCTCGA GCATCGTCGG CGGCACCAGT G        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:117:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:                             - - GGACGAATTC TTAAAGACCA GTTTTTTTGC G        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:118:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 32 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118:                             - - GAGCTCTCGA GTATCATCGG AGGTGAAGTT GC       - #                  - #              32                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:119:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119:                             - - GGACCTCGAG AATTATGCGC CGTCATTTGC         - #                  - #               30                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:120:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 806 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..736                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 656, change G to                      S.  At - # pos. aa 219, substitute Xaa."                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:                             - - C TCG CAG TCT GCA CCC TTG GTG CTA GGC GTT - #CCT GAC TTT TGG AAC             46                                                                          Ser Gln Ser Ala Pro Leu Val Leu Gly V - #al Pro Asp Phe Trp Asn                 1              - # 5                 - # 10                 - # 15         - - AGG TTA GAT GGC AGA ATC GTT GGA GGA CAC GA - #T ACT AGC ATT GAC AAA           94                                                                       Arg Leu Asp Gly Arg Ile Val Gly Gly His As - #p Thr Ser Ile Asp Lys                            20 - #                 25 - #                 30              - - CAT CCT CAT CAA GTA TCT TTG ATT TAC ACA AA - #C CAC AAT TGT GGT GGT          142                                                                       His Pro His Gln Val Ser Leu Ile Tyr Thr As - #n His Asn Cys Gly Gly                        35     - #             40     - #             45                  - - TCT TTA ATT GCC AAA AAC TGG GTT TTA ACA GC - #A GCT CAT TGC ATC AGC          190                                                                       Ser Leu Ile Ala Lys Asn Trp Val Leu Thr Al - #a Ala His Cys Ile Ser                    50         - #         55         - #         60                      - - TCA ACC TAC TAC AGA GTC CGG GTA GGA AGT TC - #A ATC AAG AAC AGT GGT          238                                                                       Ser Thr Tyr Tyr Arg Val Arg Val Gly Ser Se - #r Ile Lys Asn Ser Gly                65             - #     70             - #     75                          - - GGT GTT GTT CAC AGC GTT AAA AAC CAA ATC AA - #G CAT CCA AAA TTC GGT          286                                                                       Gly Val Val His Ser Val Lys Asn Gln Ile Ly - #s His Pro Lys Phe Gly            80                 - # 85                 - # 90                 - # 95       - - GAT TCG GCG ACA CTC GAC TTC GAT TTT GCA CT - #T CTG GAA TTG GAT GAA          334                                                                       Asp Ser Ala Thr Leu Asp Phe Asp Phe Ala Le - #u Leu Glu Leu Asp Glu                           100  - #               105  - #               110              - - CCA GTT ACA GTA ACA AAA GAC GTC AAC ATC AT - #C AAA CTA GTA GAC CAA          382                                                                       Pro Val Thr Val Thr Lys Asp Val Asn Ile Il - #e Lys Leu Val Asp Gln                       115      - #           120      - #           125                  - - GAT GTA GAA TTA ACA CCT GGA ACT ATG TGC AC - #T GTT ACT GGA TGG GGA          430                                                                       Asp Val Glu Leu Thr Pro Gly Thr Met Cys Th - #r Val Thr Gly Trp Gly                   130          - #       135          - #       140                      - - TCA ACT GGA TCT GGT GGT CCA ATT ACA AAT GT - #T CTA CAA GAA GTC GAA          478                                                                       Ser Thr Gly Ser Gly Gly Pro Ile Thr Asn Va - #l Leu Gln Glu Val Glu               145              - #   150              - #   155                          - - GTT CCA TTT ATC GAC TTC AAC ACC TGC CGA AA - #A TCC TAC TCA ACC AGC          526                                                                       Val Pro Phe Ile Asp Phe Asn Thr Cys Arg Ly - #s Ser Tyr Ser Thr Ser           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - TTA ACC GAC CGT ATG TTC TGC GCT GGA TTT TT - #G GGA ATT GGT GGT        AAG      574                                                                    Leu Thr Asp Arg Met Phe Cys Ala Gly Phe Le - #u Gly Ile Gly Gly Lys                          180  - #               185  - #               190              - - GAC GCT TGT CAA GGT GAC TCT GGT GGC CCA GT - #T GTT GTC GAT GGT GTT          622                                                                       Asp Ala Cys Gln Gly Asp Ser Gly Gly Pro Va - #l Val Val Asp Gly Val                       195      - #           200      - #           205                  - - CTT CAC GGA ATC GTA TCA TGG GGA CGT GGT TG - #C SCC CTT CCT GAC TAC          670                                                                       Leu His Gly Ile Val Ser Trp Gly Arg Gly Cy - #s Xaa Leu Pro Asp Tyr                   210          - #       215          - #       220                      - - CCC GGA GTC TAC TCT AAG ATC TCA TAT GCC CG - #T GAC TGG ATT AAG GAA          718                                                                       Pro Gly Val Tyr Ser Lys Ile Ser Tyr Ala Ar - #g Asp Trp Ile Lys Glu               225              - #   230              - #   235                          - - AAT CAC TGG TGT TTA ATT TAATATTTAT TATACCAAAT AA - #TTATATAT                 766                                                                       Asn His Trp Cys Leu Ile                                                       240                 2 - #45                                                    - - AAATATATAC TATTTTAAAT ACAAAAAAAA AAAAAAAAAA     - #                      - #   806                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:121:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 245 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:                             - - Ser Gln Ser Ala Pro Leu Val Leu Gly Val Pr - #o Asp Phe Trp Asn Arg        1               5 - #                 10 - #                 15              - - Leu Asp Gly Arg Ile Val Gly Gly His Asp Th - #r Ser Ile Asp Lys His                   20     - #             25     - #             30                  - - Pro His Gln Val Ser Leu Ile Tyr Thr Asn Hi - #s Asn Cys Gly Gly Ser               35         - #         40         - #         45                      - - Leu Ile Ala Lys Asn Trp Val Leu Thr Ala Al - #a His Cys Ile Ser Ser           50             - #     55             - #     60                          - - Thr Tyr Tyr Arg Val Arg Val Gly Ser Ser Il - #e Lys Asn Ser Gly Gly       65                 - # 70                 - # 75                 - # 80       - - Val Val His Ser Val Lys Asn Gln Ile Lys Hi - #s Pro Lys Phe Gly Asp                       85 - #                 90 - #                 95              - - Ser Ala Thr Leu Asp Phe Asp Phe Ala Leu Le - #u Glu Leu Asp Glu Pro                  100      - #           105      - #           110                  - - Val Thr Val Thr Lys Asp Val Asn Ile Ile Ly - #s Leu Val Asp Gln Asp              115          - #       120          - #       125                      - - Val Glu Leu Thr Pro Gly Thr Met Cys Thr Va - #l Thr Gly Trp Gly Ser          130              - #   135              - #   140                          - - Thr Gly Ser Gly Gly Pro Ile Thr Asn Val Le - #u Gln Glu Val Glu Val      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Pro Phe Ile Asp Phe Asn Thr Cys Arg Lys Se - #r Tyr Ser Thr Ser        Leu                                                                                             165  - #               170  - #               175             - - Thr Asp Arg Met Phe Cys Ala Gly Phe Leu Gl - #y Ile Gly Gly Lys Asp                  180      - #           185      - #           190                  - - Ala Cys Gln Gly Asp Ser Gly Gly Pro Val Va - #l Val Asp Gly Val Leu              195          - #       200          - #       205                      - - His Gly Ile Val Ser Trp Gly Arg Gly Cys Xa - #a Leu Pro Asp Tyr Pro          210              - #   215              - #   220                          - - Gly Val Tyr Ser Lys Ile Ser Tyr Ala Arg As - #p Trp Ile Lys Glu Asn      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - His Trp Cys Leu Ile                                                                      245                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:122:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 436 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..436                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 301, change A to                     W; at - #pos. bp 342, change C to Y; at pos. bp 39 - #7,                      change C - #to Y; at pos. bp 431, change G to S.  - #At       pos.                                                                                           aa 100, - #114 and 144, substitute Xaa."                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:                             - - A GTT AGT CTT TCC AAT TCG ATC AGA CCT TCT - #TGT TTA TGG GCC AAT             46                                                                         Val Ser Leu Ser Asn Ser Ile Arg Pro S - #er Cys Leu Trp Ala Asn                 1              - # 5                 - # 10                 - # 15         - - GAC GAG TTC GAC ACA GAT AGT TCA ATT GCT AC - #T GGT TGG GGA AAG ATA           94                                                                       Asp Glu Phe Asp Thr Asp Ser Ser Ile Ala Th - #r Gly Trp Gly Lys Ile                            20 - #                 25 - #                 30              - - GAC TAT GCT GAG AGC AGA AGT GAT GAC CTA CT - #G AAA GTA GTA CTG AAA          142                                                                       Asp Tyr Ala Glu Ser Arg Ser Asp Asp Leu Le - #u Lys Val Val Leu Lys                        35     - #             40     - #             45                  - - ATT ATT GAT AAT AGG CAA TGC GCT CCC TTA TA - #C GTT GAT CAG ATT AAT          190                                                                       Ile Ile Asp Asn Arg Gln Cys Ala Pro Leu Ty - #r Val Asp Gln Ile Asn                    50         - #         55         - #         60                      - - AGA AGA AGA TTG AGA AAT GGA ATT GTA GAT AC - #A CAG ATG TGT GCA GGA          238                                                                       Arg Arg Arg Leu Arg Asn Gly Ile Val Asp Th - #r Gln Met Cys Ala Gly                65             - #     70             - #     75                          - - GAA TTG GAT GGT GGC AAA GAC ACT TGC CAG GG - #A GAT TCA GGT GGG CCA          286                                                                       Glu Leu Asp Gly Gly Lys Asp Thr Cys Gln Gl - #y Asp Ser Gly Gly Pro            80                 - # 85                 - # 90                 - # 95       - - TTG CAA ATA ACA AAW CAA AGC AAC AAA TGT AT - #C TTC TAC ATA GTG GGA          334                                                                       Leu Gln Ile Thr Xaa Gln Ser Asn Lys Cys Il - #e Phe Tyr Ile Val Gly                           100  - #               105  - #               110              - - ATA ACA TYA TTC GGA AGG GGA TGT GGT GCT CC - #T AAT AGC CCC GGT GTT          382                                                                       Ile Thr Xaa Phe Gly Arg Gly Cys Gly Ala Pr - #o Asn Ser Pro Gly Val                       115      - #           120      - #           125                  - - TAT ACT AGA GTC AGY AAG TAT GTT GAC TGG AT - #T GAA AGT GTT GTT TGG          430                                                                       Tyr Thr Arg Val Ser Lys Tyr Val Asp Trp Il - #e Glu Ser Val Val Trp                   130          - #       135          - #       140                      - - SCA AAT                - #                  - #                  -      #          436                                                                  Xaa Asn                                                                           145                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:123:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 145 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:                             - - Val Ser Leu Ser Asn Ser Ile Arg Pro Ser Cy - #s Leu Trp Ala Asn        Asp                                                                               1               5 - #                 10 - #                 15             - - Glu Phe Asp Thr Asp Ser Ser Ile Ala Thr Gl - #y Trp Gly Lys Ile Asp                   20     - #             25     - #             30                  - - Tyr Ala Glu Ser Arg Ser Asp Asp Leu Leu Ly - #s Val Val Leu Lys Ile               35         - #         40         - #         45                      - - Ile Asp Asn Arg Gln Cys Ala Pro Leu Tyr Va - #l Asp Gln Ile Asn Arg           50             - #     55             - #     60                          - - Arg Arg Leu Arg Asn Gly Ile Val Asp Thr Gl - #n Met Cys Ala Gly Glu       65                 - # 70                 - # 75                 - # 80       - - Leu Asp Gly Gly Lys Asp Thr Cys Gln Gly As - #p Ser Gly Gly Pro Leu                       85 - #                 90 - #                 95              - - Gln Ile Thr Xaa Gln Ser Asn Lys Cys Ile Ph - #e Tyr Ile Val Gly Ile                  100      - #           105      - #           110                  - - Thr Xaa Phe Gly Arg Gly Cys Gly Ala Pro As - #n Ser Pro Gly Val Tyr              115          - #       120          - #       125                      - - Thr Arg Val Ser Lys Tyr Val Asp Trp Ile Gl - #u Ser Val Val Trp Xaa          130              - #   135              - #   140                          - - Asn                                                                      145                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:124:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 307 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..306                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:                             - - CAA ATA TTT AAG CAT GAA CCT GTG AAC GTG GT - #A TTA TTT CTT GTA GAA           48                                                                       Gln Ile Phe Lys His Glu Pro Val Asn Val Va - #l Leu Phe Leu Val Glu             1               5 - #                 10 - #                 15              - - GAT CGC TTC AAT TTT GAA ATT ACT AAC GCG CG - #C CCT GTT GCC CTA CCA           96                                                                       Asp Arg Phe Asn Phe Glu Ile Thr Asn Ala Ar - #g Pro Val Ala Leu Pro                        20     - #             25     - #             30                  - - GCA GAG AAC GAG GAA ACC GAA ACA GGG TCA CC - #A CTC ACG GTG ACG GGT          144                                                                       Ala Glu Asn Glu Glu Thr Glu Thr Gly Ser Pr - #o Leu Thr Val Thr Gly                    35         - #         40         - #         45                      - - TGG GGA ACT ACA GAG AGT ACT GAA TCA TCA CA - #C CAC CTG AAA GAA GTT          192                                                                       Trp Gly Thr Thr Glu Ser Thr Glu Ser Ser Hi - #s His Leu Lys Glu Val                50             - #     55             - #     60                          - - GAA GTG AAC GCT GTA TCT AAT AGT GAA TGT CA - #A AAG GCC TAT GAA GAT          240                                                                       Glu Val Asn Ala Val Ser Asn Ser Glu Cys Gl - #n Lys Ala Tyr Glu Asp            65                 - # 70                 - # 75                 - # 80       - - CTT GCT ACT ATA TCA TCA CAT GAG ATA TGT GC - #A AGC GTT CCT GGT GGC          288                                                                       Leu Ala Thr Ile Ser Ser His Glu Ile Cys Al - #a Ser Val Pro Gly Gly                            85 - #                 90 - #                 95              - - GGC AAA GAT TCT TGT CAA G        - #                  - #                      - #307                                                                  Gly Lys Asp Ser Cys Gln                                                                   100                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:125:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 102 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:                             - - Gln Ile Phe Lys His Glu Pro Val Asn Val Va - #l Leu Phe Leu Val Glu        1               5 - #                 10 - #                 15              - - Asp Arg Phe Asn Phe Glu Ile Thr Asn Ala Ar - #g Pro Val Ala Leu Pro                   20     - #             25     - #             30                  - - Ala Glu Asn Glu Glu Thr Glu Thr Gly Ser Pr - #o Leu Thr Val Thr Gly               35         - #         40         - #         45                      - - Trp Gly Thr Thr Glu Ser Thr Glu Ser Ser Hi - #s His Leu Lys Glu Val           50             - #     55             - #     60                          - - Glu Val Asn Ala Val Ser Asn Ser Glu Cys Gl - #n Lys Ala Tyr Glu Asp       65                 - # 70                 - # 75                 - # 80       - - Leu Ala Thr Ile Ser Ser His Glu Ile Cys Al - #a Ser Val Pro Gly Gly                       85 - #                 90 - #                 95              - - Gly Lys Asp Ser Cys Gln                                                              100                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:126:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 758 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..738                                                          (D) OTHER INFORMATION: - #/note= "At pos. aa 241, substitute                       Xaa."                                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:                             - - AAG GTA CTG ATC GTT TTA GCA GTC ATT GAA TT - #C GCA TCA GCG TCT TCA           48                                                                       Lys Val Leu Ile Val Leu Ala Val Ile Glu Ph - #e Ala Ser Ala Ser Ser             1               5 - #                 10 - #                 15              - - ATC GGC TGG AGA ATC GTG GGT GGT GAA AAT GC - #T AAA GAA AAA TCG GTG           96                                                                       Ile Gly Trp Arg Ile Val Gly Gly Glu Asn Al - #a Lys Glu Lys Ser Val                        20     - #             25     - #             30                  - - CCC TAT CAA GTT TCA CTT CGA AAT GCT GAA AA - #C AAA CAT TTC TGT GGA          144                                                                       Pro Tyr Gln Val Ser Leu Arg Asn Ala Glu As - #n Lys His Phe Cys Gly                    35         - #         40         - #         45                      - - GGA GCA ATT ATT GAC GAT TAT TGG GTT TTG AC - #T GCT GCT CAT TGC ATG          192                                                                       Gly Ala Ile Ile Asp Asp Tyr Trp Val Leu Th - #r Ala Ala His Cys Met                50             - #     55             - #     60                          - - GGA CAA CGT TTT GAA GTC GTT GCC GGC GTG AA - #T AAA CTG GAT GAA GTA          240                                                                       Gly Gln Arg Phe Glu Val Val Ala Gly Val As - #n Lys Leu Asp Glu Val            65                 - # 70                 - # 75                 - # 80       - - GGT GAA CGA TAT AGA ATA GAA AAA ACT ATT AC - #T GAC AAG TTT GAT GAA          288                                                                       Gly Glu Arg Tyr Arg Ile Glu Lys Thr Ile Th - #r Asp Lys Phe Asp Glu                            85 - #                 90 - #                 95              - - CAA ACT GCC GCA AAT GAT TTG GCA CTT GTA AA - #A CTT CGG AAT AAA ATA          336                                                                       Gln Thr Ala Ala Asn Asp Leu Ala Leu Val Ly - #s Leu Arg Asn Lys Ile                       100      - #           105      - #           110                  - - AAA TTC AGC GAT AAA GTG CAA AAA ATT CAA TT - #T GAA GAT AAA TAT ATC          384                                                                       Lys Phe Ser Asp Lys Val Gln Lys Ile Gln Ph - #e Glu Asp Lys Tyr Ile                   115          - #       120          - #       125                      - - GGA GGC GGA GAG GAT GCT CGT TTG ACT GGA TG - #G GGA CGA TTG GGA AAA          432                                                                       Gly Gly Gly Glu Asp Ala Arg Leu Thr Gly Tr - #p Gly Arg Leu Gly Lys               130              - #   135              - #   140                          - - GAT TCA CCG CCA CCT AAT GAT TTA CAG GAA TT - #A AAT ACA TTT ACC ATC          480                                                                       Asp Ser Pro Pro Pro Asn Asp Leu Gln Glu Le - #u Asn Thr Phe Thr Ile           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - CCC CAA AGT GTT TGC AGA AGA ATG TTT AAT GA - #G GAT AAG ATT CCA        ATC      528                                                                    Pro Gln Ser Val Cys Arg Arg Met Phe Asn Gl - #u Asp Lys Ile Pro Ile                          165  - #               170  - #               175              - - CAC GAT AGC CAA ATA TGC ACT TTT GCA GAT AT - #G GGC AAG GGT GCA TGT          576                                                                       His Asp Ser Gln Ile Cys Thr Phe Ala Asp Me - #t Gly Lys Gly Ala Cys                       180      - #           185      - #           190                  - - AAG GGT GAT TCC GGT GGC CCC TTA GTC ATC AA - #T GGA CAA CTT CAT GGA          624                                                                       Lys Gly Asp Ser Gly Gly Pro Leu Val Ile As - #n Gly Gln Leu His Gly                   195          - #       200          - #       205                      - - ATT GTT TCC TGG GGC ATT CCT TGC GCT GTC GG - #C AAG CCT GAT GTA TTC          672                                                                       Ile Val Ser Trp Gly Ile Pro Cys Ala Val Gl - #y Lys Pro Asp Val Phe               210              - #   215              - #   220                          - - ACA AGA GTT TCT CAT TAT GTC GAT TGG ATT AA - #A TCC AAA ATT GCC AAA          720                                                                       Thr Arg Val Ser His Tyr Val Asp Trp Ile Ly - #s Ser Lys Ile Ala Lys           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - TAA AAT TGT TTA GTG TAT TAAAAAAAAA AAAAAAAAAA  - #                      - #    758                                                                    Xaa Asn Cys Leu Val Tyr                                                                       245                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:127:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 246 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127:                             - - Lys Val Leu Ile Val Leu Ala Val Ile Glu Ph - #e Ala Ser Ala Ser Ser        1               5 - #                 10 - #                 15              - - Ile Gly Trp Arg Ile Val Gly Gly Glu Asn Al - #a Lys Glu Lys Ser Val                   20     - #             25     - #             30                  - - Pro Tyr Gln Val Ser Leu Arg Asn Ala Glu As - #n Lys His Phe Cys Gly               35         - #         40         - #         45                      - - Gly Ala Ile Ile Asp Asp Tyr Trp Val Leu Th - #r Ala Ala His Cys Met           50             - #     55             - #     60                          - - Gly Gln Arg Phe Glu Val Val Ala Gly Val As - #n Lys Leu Asp Glu Val       65                 - # 70                 - # 75                 - # 80       - - Gly Glu Arg Tyr Arg Ile Glu Lys Thr Ile Th - #r Asp Lys Phe Asp Glu                       85 - #                 90 - #                 95              - - Gln Thr Ala Ala Asn Asp Leu Ala Leu Val Ly - #s Leu Arg Asn Lys Ile                  100      - #           105      - #           110                  - - Lys Phe Ser Asp Lys Val Gln Lys Ile Gln Ph - #e Glu Asp Lys Tyr Ile              115          - #       120          - #       125                      - - Gly Gly Gly Glu Asp Ala Arg Leu Thr Gly Tr - #p Gly Arg Leu Gly Lys          130              - #   135              - #   140                          - - Asp Ser Pro Pro Pro Asn Asp Leu Gln Glu Le - #u Asn Thr Phe Thr Ile      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Pro Gln Ser Val Cys Arg Arg Met Phe Asn Gl - #u Asp Lys Ile Pro        Ile                                                                                             165  - #               170  - #               175             - - His Asp Ser Gln Ile Cys Thr Phe Ala Asp Me - #t Gly Lys Gly Ala Cys                  180      - #           185      - #           190                  - - Lys Gly Asp Ser Gly Gly Pro Leu Val Ile As - #n Gly Gln Leu His Gly              195          - #       200          - #       205                      - - Ile Val Ser Trp Gly Ile Pro Cys Ala Val Gl - #y Lys Pro Asp Val Phe          210              - #   215              - #   220                          - - Thr Arg Val Ser His Tyr Val Asp Trp Ile Ly - #s Ser Lys Ile Ala Lys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Xaa Asn Cys Leu Val Tyr                                                                  245                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:128:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 815 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..762                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 453, change A to                     M; at - #454, change G to V; at 456, G to V; at - #457, A     to                                                                                            M; at - #460, A to R; at 470, G to S; at 493, A - # to R.      At                                                                                           pos. aa - #120, 136, 152, 153, 154, 157 and 165,                              substitute - #Xaa."                                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128:                             - - ATT TTA TTA AGC GCA TTA TTT GCA AGT GTA AT - #T TGC TCC TTT AAC GCG           48                                                                       Ile Leu Leu Ser Ala Leu Phe Ala Ser Val Il - #e Cys Ser Phe Asn Ala             1               5 - #                 10 - #                 15              - - GAA GTA CAA AAT CGA ATC GTT GGT GGC AAT GA - #T GTA AGT ATT TCA AAA           96                                                                       Glu Val Gln Asn Arg Ile Val Gly Gly Asn As - #p Val Ser Ile Ser Lys                        20     - #             25     - #             30                  - - ATT GGG TGG CAA GTA TCT ATT CAA AGT AAT AA - #C CAA CAT TTC TGT GGT          144                                                                       Ile Gly Trp Gln Val Ser Ile Gln Ser Asn As - #n Gln His Phe Cys Gly                    35         - #         40         - #         45                      - - GGT TCA ATC ATT GCT AAA GAT TGG GTA CTG AC - #T TCT TCT CAA TGC GTC          192                                                                       Gly Ser Ile Ile Ala Lys Asp Trp Val Leu Th - #r Ser Ser Gln Cys Val                50             - #     55             - #     60                          - - GTG GAC AAA CAA AGT CCA CCG AAG GAT TTA AC - #T GTT CGT GTT GGA ACT          240                                                                       Val Asp Lys Gln Ser Pro Pro Lys Asp Leu Th - #r Val Arg Val Gly Thr            65                 - # 70                 - # 75                 - # 80       - - AGC ACT CAC AAT GAT GGA GGA AAA GTG TAT GA - #T GTT ATT GAA ATT ATA          288                                                                       Ser Thr His Asn Asp Gly Gly Lys Val Tyr As - #p Val Ile Glu Ile Ile                            85 - #                 90 - #                 95              - - AAA CAT CCG AAA TAT AAT AAA GCA GTG CCA GA - #T GAT TTT GAT GTT GCA          336                                                                       Lys His Pro Lys Tyr Asn Lys Ala Val Pro As - #p Asp Phe Asp Val Ala                       100      - #           105      - #           110                  - - CTT TTA CGG ATC AAA GAG CCA ATC ATT TAC TC - #C ATG CAC AGT AAC TCC          384                                                                       Leu Leu Arg Ile Lys Glu Pro Xaa Ile Tyr Se - #r Met His Ser Asn Ser                   115          - #       120          - #       125                      - - TGT AAA ATT AAT ACA ATC GGG AAA GAA GTA CC - #G AAG GGA ACA ACT TTG          432                                                                       Cys Lys Ile Asn Thr Ile Gly Xaa Glu Val Pr - #o Lys Gly Thr Thr Leu               130              - #   135              - #   140                          - - AGT GTA ACT GGA TGG GGC GCM VCV MAG RAA TG - #G GGG CSC AAT TTC GCC          480                                                                       Ser Val Thr Gly Trp Gly Ala Xaa Xaa Xaa Tr - #p Gly Xaa Asn Phe Ala           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - AAA GTT ACA AGA RAG TTA AAG TTA AAG CTA CT - #C AAG TCA AGA ATG        CAA      528                                                                    Lys Val Thr Arg Xaa Leu Lys Leu Lys Leu Le - #u Lys Ser Arg Met Gln                          165  - #               170  - #               175              - - GAA CAG TCT GCT ATT AAC AGT GAC ATC ATT TC - #T GAC AGT ATG ATG TGC          576                                                                       Glu Gln Ser Ala Ile Asn Ser Asp Ile Ile Se - #r Asp Ser Met Met Cys                       180      - #           185      - #           190                  - - GCT GGT TTT CCT CAA GGA CAA AAA GAT ACT TG - #T CAT GGG GAT AGC GGT          624                                                                       Ala Gly Phe Pro Gln Gly Gln Lys Asp Thr Cy - #s His Gly Asp Ser Gly                   195          - #       200          - #       205                      - - GGC ACT GTA GAT AAA AAA CAG GTT CAA GTA GG - #A GTT ATA TCC TGG AGG          672                                                                       Gly Thr Val Asp Lys Lys Gln Val Gln Val Gl - #y Val Ile Ser Trp Arg               210              - #   215              - #   220                          - - CGA GGA TGC GCG CGA CCT GGA TAT CCT GGC GT - #A TAT ACA AAA TTG AGC          720                                                                       Arg Gly Cys Ala Arg Pro Gly Tyr Pro Gly Va - #l Tyr Thr Lys Leu Ser           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - CAC CCG GAA ATC CAA CAG TTT ATT AAA AAC AA - #T GTA AAA CTT                 - # 762                                                                   His Pro Glu Ile Gln Gln Phe Ile Lys Asn As - #n Val Lys Leu                                   245  - #               250                                     - - TAAATCATAA AACTGTATGA AAATAACAAT AACAATTACG GGAAAAAAAA AA - #A               815                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:129:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 254 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129:                             - - Ile Leu Leu Ser Ala Leu Phe Ala Ser Val Il - #e Cys Ser Phe Asn Ala        1               5 - #                 10 - #                 15              - - Glu Val Gln Asn Arg Ile Val Gly Gly Asn As - #p Val Ser Ile Ser Lys                   20     - #             25     - #             30                  - - Ile Gly Trp Gln Val Ser Ile Gln Ser Asn As - #n Gln His Phe Cys Gly               35         - #         40         - #         45                      - - Gly Ser Ile Ile Ala Lys Asp Trp Val Leu Th - #r Ser Ser Gln Cys Val           50             - #     55             - #     60                          - - Val Asp Lys Gln Ser Pro Pro Lys Asp Leu Th - #r Val Arg Val Gly Thr       65                 - # 70                 - # 75                 - # 80       - - Ser Thr His Asn Asp Gly Gly Lys Val Tyr As - #p Val Ile Glu Ile Ile                       85 - #                 90 - #                 95              - - Lys His Pro Lys Tyr Asn Lys Ala Val Pro As - #p Asp Phe Asp Val Ala                  100      - #           105      - #           110                  - - Leu Leu Arg Ile Lys Glu Pro Xaa Ile Tyr Se - #r Met His Ser Asn Ser              115          - #       120          - #       125                      - - Cys Lys Ile Asn Thr Ile Gly Xaa Glu Val Pr - #o Lys Gly Thr Thr Leu          130              - #   135              - #   140                          - - Ser Val Thr Gly Trp Gly Ala Xaa Xaa Xaa Tr - #p Gly Xaa Asn Phe Ala      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Lys Val Thr Arg Xaa Leu Lys Leu Lys Leu Le - #u Lys Ser Arg Met        Gln                                                                                             165  - #               170  - #               175             - - Glu Gln Ser Ala Ile Asn Ser Asp Ile Ile Se - #r Asp Ser Met Met Cys                  180      - #           185      - #           190                  - - Ala Gly Phe Pro Gln Gly Gln Lys Asp Thr Cy - #s His Gly Asp Ser Gly              195          - #       200          - #       205                      - - Gly Thr Val Asp Lys Lys Gln Val Gln Val Gl - #y Val Ile Ser Trp Arg          210              - #   215              - #   220                          - - Arg Gly Cys Ala Arg Pro Gly Tyr Pro Gly Va - #l Tyr Thr Lys Leu Ser      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - His Pro Glu Ile Gln Gln Phe Ile Lys Asn As - #n Val Lys Leu                             245  - #               250                                     - -  - - (2) INFORMATION FOR SEQ ID NO:130:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 855 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..759                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 693, change C to                      N.  At - # pos. aa 231, substitute Xaa."                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:                             - - ATG GCT TAT ATT ATA TTA GTT ACT TTA ATT AG - #T TTG GGA TCG TTG GTT           48                                                                       Met Ala Tyr Ile Ile Leu Val Thr Leu Ile Se - #r Leu Gly Ser Leu Val             1               5 - #                 10 - #                 15              - - TCT TCC GAA TAC CTT TCG TTT TCT ACT GAT CC - #T CGG ATA ATT GGT GGT           96                                                                       Ser Ser Glu Tyr Leu Ser Phe Ser Thr Asp Pr - #o Arg Ile Ile Gly Gly                        20     - #             25     - #             30                  - - GAA GAT GCT CCC GAG GGT TCT GCA CCA TAT CA - #G GTT TCA TTA AGA AAT          144                                                                       Glu Asp Ala Pro Glu Gly Ser Ala Pro Tyr Gl - #n Val Ser Leu Arg Asn                    35         - #         40         - #         45                      - - TCT GAC TTG CAG CAT TTT TGT GGT GGT TCC AT - #C CTA AAC AAA CGA TGG          192                                                                       Ser Asp Leu Gln His Phe Cys Gly Gly Ser Il - #e Leu Asn Lys Arg Trp                50             - #     55             - #     60                          - - ATT TTA ACA GCA GCA CAT TGT CTC GAA CCT GG - #T TTT TTA AAT TCT GTA          240                                                                       Ile Leu Thr Ala Ala His Cys Leu Glu Pro Gl - #y Phe Leu Asn Ser Val            65                 - # 70                 - # 75                 - # 80       - - TAC ATG GGT TCG AAT TTG TTG GAT CGA AAA GG - #C AGA TAT TAC GAT GTA          288                                                                       Tyr Met Gly Ser Asn Leu Leu Asp Arg Lys Gl - #y Arg Tyr Tyr Asp Val                            85 - #                 90 - #                 95              - - GAA AGA TTT GTG ATG CAC CAT AAT TAT ACT GG - #A AAG ATA GTT GCC AAT          336                                                                       Glu Arg Phe Val Met His His Asn Tyr Thr Gl - #y Lys Ile Val Ala Asn                       100      - #           105      - #           110                  - - GTC GCT GAT ATA GGT CTA ATA AAA CTA GCA GA - #A GAT ATA AAA TTC AGT          384                                                                       Val Ala Asp Ile Gly Leu Ile Lys Leu Ala Gl - #u Asp Ile Lys Phe Ser                   115          - #       120          - #       125                      - - GAC AAG GTA CAA CCT GTA AAA ATT CAT CAA AC - #T CAA ATC AAG GGC GGA          432                                                                       Asp Lys Val Gln Pro Val Lys Ile His Gln Th - #r Gln Ile Lys Gly Gly               130              - #   135              - #   140                          - - GAG ATT TGC AAA GCT ACT GGA TGG GGC AGG TT - #G GGT GCT GAT CAG CCT          480                                                                       Glu Ile Cys Lys Ala Thr Gly Trp Gly Arg Le - #u Gly Ala Asp Gln Pro           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - GTA CCA AAT AAA TTA CAA CAA TTG GAG ACA AT - #T GCT ATT AGT GAT        GAG      528                                                                    Val Pro Asn Lys Leu Gln Gln Leu Glu Thr Il - #e Ala Ile Ser Asp Glu                          165  - #               170  - #               175              - - AAA TGT TAT GCA GAT ACA GGG TTT TTA GAA CC - #T ACA TCT CAA ATA TGT          576                                                                       Lys Cys Tyr Ala Asp Thr Gly Phe Leu Glu Pr - #o Thr Ser Gln Ile Cys                       180      - #           185      - #           190                  - - GTA TTC AGT GCA TTT GGA AAA GGA GTT TGT TT - #T GGA GAT TCT GGT GGT          624                                                                       Val Phe Ser Ala Phe Gly Lys Gly Val Cys Ph - #e Gly Asp Ser Gly Gly                   195          - #       200          - #       205                      - - CCA TTA GTT TAC AAA GGT GAA CAA GTA GGA GT - #T GCA TCA TTC ATC ATG          672                                                                       Pro Leu Val Tyr Lys Gly Glu Gln Val Gly Va - #l Ala Ser Phe Ile Met               210              - #   215              - #   220                          - - ATC ACT TGT GGT GGT GGC AGN CCA GAT GTA TT - #T GTT AGA GTA CTC GAT          720                                                                       Ile Thr Cys Gly Gly Gly Xaa Pro Asp Val Ph - #e Val Arg Val Leu Asp           225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - TAT CAG GAT TGG ATA AAT TCA TTT ATT TCT GG - #A GAT AAC TAGTCTTTAA           769                                                                      Tyr Gln Asp Trp Ile Asn Ser Phe Ile Ser Gl - #y Asp Asn                                       245  - #               250                                     - - TGTAAAATGA ACTATTATAA TATATATTTT TTATTCTTAT AAAATATATA CA -             #TTTTATTA    829                                                                 - - CGCACAAAAA AAAAAAAAAA AAAAAA          - #                  - #                 855                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:131:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 253 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:                             - - Met Ala Tyr Ile Ile Leu Val Thr Leu Ile Se - #r Leu Gly Ser Leu Val        1               5 - #                 10 - #                 15              - - Ser Ser Glu Tyr Leu Ser Phe Ser Thr Asp Pr - #o Arg Ile Ile Gly Gly                   20     - #             25     - #             30                  - - Glu Asp Ala Pro Glu Gly Ser Ala Pro Tyr Gl - #n Val Ser Leu Arg Asn               35         - #         40         - #         45                      - - Ser Asp Leu Gln His Phe Cys Gly Gly Ser Il - #e Leu Asn Lys Arg Trp           50             - #     55             - #     60                          - - Ile Leu Thr Ala Ala His Cys Leu Glu Pro Gl - #y Phe Leu Asn Ser Val       65                 - # 70                 - # 75                 - # 80       - - Tyr Met Gly Ser Asn Leu Leu Asp Arg Lys Gl - #y Arg Tyr Tyr Asp Val                       85 - #                 90 - #                 95              - - Glu Arg Phe Val Met His His Asn Tyr Thr Gl - #y Lys Ile Val Ala Asn                  100      - #           105      - #           110                  - - Val Ala Asp Ile Gly Leu Ile Lys Leu Ala Gl - #u Asp Ile Lys Phe Ser              115          - #       120          - #       125                      - - Asp Lys Val Gln Pro Val Lys Ile His Gln Th - #r Gln Ile Lys Gly Gly          130              - #   135              - #   140                          - - Glu Ile Cys Lys Ala Thr Gly Trp Gly Arg Le - #u Gly Ala Asp Gln Pro      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Val Pro Asn Lys Leu Gln Gln Leu Glu Thr Il - #e Ala Ile Ser Asp        Glu                                                                                             165  - #               170  - #               175             - - Lys Cys Tyr Ala Asp Thr Gly Phe Leu Glu Pr - #o Thr Ser Gln Ile Cys                  180      - #           185      - #           190                  - - Val Phe Ser Ala Phe Gly Lys Gly Val Cys Ph - #e Gly Asp Ser Gly Gly              195          - #       200          - #       205                      - - Pro Leu Val Tyr Lys Gly Glu Gln Val Gly Va - #l Ala Ser Phe Ile Met          210              - #   215              - #   220                          - - Ile Thr Cys Gly Gly Gly Xaa Pro Asp Val Ph - #e Val Arg Val Leu Asp      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Tyr Gln Asp Trp Ile Asn Ser Phe Ile Ser Gl - #y Asp Asn                                  245  - #               250                                     - -  - - (2) INFORMATION FOR SEQ ID NO:132:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 595 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..595                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 509, change C to                     Y; at - #pos. 556, change C to Y; at pos. 557, change - #     C to                                                                                          Y; at - #561, change C to Y; at 573, change C to Y - #.       At                                                                                             pos. aa. - #170, 186, 187, 190 and 191, substitute Xaa."       - -         (xi) SEQUENCE DESCRIPTION: SEQ - #ID NO:132:                      - - T GGT TCC ATC TTG AAC AAA CGT TGG ATT GTA - #ACA GCT GCA CAT TGC             46                                                                         - -   Gly Ser Ile Leu Asn Lys Arg Trp Ile V - #al Thr Ala Ala His Cys            1              - # 5                 - # 10                 - # 15         - - CTA AAC GCT GGC ATT TTA AAA TCC GTC TAT TT - #G GGA TCA AAC TCA TTA           94                                                                       Leu Asn Ala Gly Ile Leu Lys Ser Val Tyr Le - #u Gly Ser Asn Ser Leu                            20 - #                 25 - #                 30              - - GAT GGC GAT GGT ACA TAC TAC GAC GTC GAA CG - #T TTT GTG ATG CAT GAT          142                                                                       Asp Gly Asp Gly Thr Tyr Tyr Asp Val Glu Ar - #g Phe Val Met His Asp                        35     - #             40     - #             45                  - - AAA TAT ACA CCA AGA ATC ACT GTC AAC TAT GC - #T GAT ATT GGT CTA ATA          190                                                                       Lys Tyr Thr Pro Arg Ile Thr Val Asn Tyr Al - #a Asp Ile Gly Leu Ile                    50         - #         55         - #         60                      - - AAA GTG GCA AAA GAC ATT GTA TTC GGT GAC AA - #A GTC CAA CCG ATC AAA          238                                                                       Lys Val Ala Lys Asp Ile Val Phe Gly Asp Ly - #s Val Gln Pro Ile Lys                65             - #     70             - #     75                          - - ATT AGC AAG AGA AAC ATC AAG GGT GGT GAA AT - #T TGC AAG GCA ACT GGT          286                                                                       Ile Ser Lys Arg Asn Ile Lys Gly Gly Glu Il - #e Cys Lys Ala Thr Gly            80                 - # 85                 - # 90                 - # 95       - - TGG GGT CTA TTA GGT TCT GTG GAC TCA GTA CC - #A AAC GAA TTA CAA CAA          334                                                                       Trp Gly Leu Leu Gly Ser Val Asp Ser Val Pr - #o Asn Glu Leu Gln Gln                           100  - #               105  - #               110              - - GTA GAA ACC ACT GCA ATA ACA GAC GAA AAG TG - #C TTT GAA TTG ACT CAA          382                                                                       Val Glu Thr Thr Ala Ile Thr Asp Glu Lys Cy - #s Phe Glu Leu Thr Gln                       115      - #           120      - #           125                  - - TTC ATT GAC CCA ACT TCG CAA ATA TGT ACA TT - #C AGG GAA TTT GGT AGA          430                                                                       Phe Ile Asp Pro Thr Ser Gln Ile Cys Thr Ph - #e Arg Glu Phe Gly Arg                   130          - #       135          - #       140                      - - GGC ATT TGC TTT GGT GAT TCT GGT GGA CCA CT - #A GTT TAC AAA AAT GAA          478                                                                       Gly Ile Cys Phe Gly Asp Ser Gly Gly Pro Le - #u Val Tyr Lys Asn Glu               145              - #   150              - #   155                          - - CTT GTT GGC ATT ACA TCG ATG CAC TTA TAC YC - #C TGC AGA GGT GGC AGG          526                                                                       Leu Val Gly Ile Thr Ser Met His Leu Tyr Xa - #a Cys Arg Gly Gly Arg           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - CCA GAT ATT TTT TGT GAA AGT GCG AGA TTY YC - #A AYC CTG GAT TAA        AYT      574                                                                    Pro Asp Ile Phe Cys Glu Ser Ala Arg Phe Xa - #a Xaa Leu Asp Xaa Xaa                          180  - #               185  - #               190              - - CTG AAA TTG AAA AAA ATT AAA       - #                  - #                     595                                                                     Leu Lys Leu Lys Lys Ile Lys                                                               195                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:133:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 198 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:                             - - Gly Ser Ile Leu Asn Lys Arg Trp Ile Val Th - #r Ala Ala His Cys Leu        1               5 - #                 10 - #                 15              - - Asn Ala Gly Ile Leu Lys Ser Val Tyr Leu Gl - #y Ser Asn Ser Leu Asp                   20     - #             25     - #             30                  - - Gly Asp Gly Thr Tyr Tyr Asp Val Glu Arg Ph - #e Val Met His Asp Lys               35         - #         40         - #         45                      - - Tyr Thr Pro Arg Ile Thr Val Asn Tyr Ala As - #p Ile Gly Leu Ile Lys           50             - #     55             - #     60                          - - Val Ala Lys Asp Ile Val Phe Gly Asp Lys Va - #l Gln Pro Ile Lys Ile       65                 - # 70                 - # 75                 - # 80       - - Ser Lys Arg Asn Ile Lys Gly Gly Glu Ile Cy - #s Lys Ala Thr Gly Trp                       85 - #                 90 - #                 95              - - Gly Leu Leu Gly Ser Val Asp Ser Val Pro As - #n Glu Leu Gln Gln Val                  100      - #           105      - #           110                  - - Glu Thr Thr Ala Ile Thr Asp Glu Lys Cys Ph - #e Glu Leu Thr Gln Phe              115          - #       120          - #       125                      - - Ile Asp Pro Thr Ser Gln Ile Cys Thr Phe Ar - #g Glu Phe Gly Arg Gly          130              - #   135              - #   140                          - - Ile Cys Phe Gly Asp Ser Gly Gly Pro Leu Va - #l Tyr Lys Asn Glu Leu      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Val Gly Ile Thr Ser Met His Leu Tyr Xaa Cy - #s Arg Gly Gly Arg        Pro                                                                                             165  - #               170  - #               175             - - Asp Ile Phe Cys Glu Ser Ala Arg Phe Xaa Xa - #a Leu Asp Xaa Xaa Leu                  180      - #           185      - #           190                  - - Lys Leu Lys Lys Ile Lys                                                          195                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:134:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 423 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..423                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134:                             - - GTA GTT ACA GCT GCA CAT TGT GTC ACC GTT GG - #A GGA CAT AAC CAA GTC           48                                                                       Val Val Thr Ala Ala His Cys Val Thr Val Gl - #y Gly His Asn Gln Val             1               5 - #                 10 - #                 15              - - GTA GCT GTT GTA GGA ACC AAC AAA TTG AGC TC - #C GGA GGC ACC ACA TAC           96                                                                       Val Ala Val Val Gly Thr Asn Lys Leu Ser Se - #r Gly Gly Thr Thr Tyr                        20     - #             25     - #             30                  - - AAA GCT GAA CGT GTT GTT GTA CAC GAA CGT TA - #T GGC AAT GCT GAT ATT          144                                                                       Lys Ala Glu Arg Val Val Val His Glu Arg Ty - #r Gly Asn Ala Asp Ile                    35         - #         40         - #         45                      - - GAC AAC GAT CTT GCC TTG ATC AAG TTG ACC CA - #A GAT GTC GTA TTC ACT          192                                                                       Asp Asn Asp Leu Ala Leu Ile Lys Leu Thr Gl - #n Asp Val Val Phe Thr                50             - #     55             - #     60                          - - GAC CGC GTA CAG CCC GTC ACC GTA TCC AGA AC - #T ACA GTC AAA GGA GGA          240                                                                       Asp Arg Val Gln Pro Val Thr Val Ser Arg Th - #r Thr Val Lys Gly Gly            65                 - # 70                 - # 75                 - # 80       - - GAA ACC TTG AGA ATC ACT GGA TGG GGT TAC AC - #C AAC CAC GGT GGC CCA          288                                                                       Glu Thr Leu Arg Ile Thr Gly Trp Gly Tyr Th - #r Asn His Gly Gly Pro                            85 - #                 90 - #                 95              - - GTT CTG CCC GAC AGT TTG CAA GAA CTT CAT GT - #A ACC GCC CAG ACC CCA          336                                                                       Val Leu Pro Asp Ser Leu Gln Glu Leu His Va - #l Thr Ala Gln Thr Pro                       100      - #           105      - #           110                  - - AGC ACA TGC CAA AAA TAC ACA CCA GCC GCC AC - #A CAA TTG TGC ACT TTC          384                                                                       Ser Thr Cys Gln Lys Tyr Thr Pro Ala Ala Th - #r Gln Leu Cys Thr Phe                   115          - #       120          - #       125                      - - TTG AAG ACT GGA CAA GGA GTT TGC AAC GGT GA - #T TCT GGA                  - #    423                                                                    Leu Lys Thr Gly Gln Gly Val Cys Asn Gly As - #p Ser Gly                           130              - #   135              - #   140                          - -  - - (2) INFORMATION FOR SEQ ID NO:135:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 141 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:                             - - Val Val Thr Ala Ala His Cys Val Thr Val Gl - #y Gly His Asn Gln Val        1               5 - #                 10 - #                 15              - - Val Ala Val Val Gly Thr Asn Lys Leu Ser Se - #r Gly Gly Thr Thr Tyr                   20     - #             25     - #             30                  - - Lys Ala Glu Arg Val Val Val His Glu Arg Ty - #r Gly Asn Ala Asp Ile               35         - #         40         - #         45                      - - Asp Asn Asp Leu Ala Leu Ile Lys Leu Thr Gl - #n Asp Val Val Phe Thr           50             - #     55             - #     60                          - - Asp Arg Val Gln Pro Val Thr Val Ser Arg Th - #r Thr Val Lys Gly Gly       65                 - # 70                 - # 75                 - # 80       - - Glu Thr Leu Arg Ile Thr Gly Trp Gly Tyr Th - #r Asn His Gly Gly Pro                       85 - #                 90 - #                 95              - - Val Leu Pro Asp Ser Leu Gln Glu Leu His Va - #l Thr Ala Gln Thr Pro                  100      - #           105      - #           110                  - - Ser Thr Cys Gln Lys Tyr Thr Pro Ala Ala Th - #r Gln Leu Cys Thr Phe              115          - #       120          - #       125                      - - Leu Lys Thr Gly Gln Gly Val Cys Asn Gly As - #p Ser Gly                      130              - #   135              - #   140                          - -  - - (2) INFORMATION FOR SEQ ID NO:136:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 410 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..408                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136:                             - - TGG GTT GTT ACT GCT GCT CAT TGT TTG AGA GG - #C AAA GAC CAC CTC CTG           48                                                                       Trp Val Val Thr Ala Ala His Cys Leu Arg Gl - #y Lys Asp His Leu Leu             1               5 - #                 10 - #                 15              - - GAC AAA CTG TTC ATT GCA GTC GGC CTG ACA AA - #T TTA GGT GAA GGA GGC           96                                                                       Asp Lys Leu Phe Ile Ala Val Gly Leu Thr As - #n Leu Gly Glu Gly Gly                        20     - #             25     - #             30                  - - ACC GTG TAT CCT GTA GAA AAA GGC ATC ATG CA - #C GAA GAA TAT GAA CAT          144                                                                       Thr Val Tyr Pro Val Glu Lys Gly Ile Met Hi - #s Glu Glu Tyr Glu His                    35         - #         40         - #         45                      - - TAT GAC ATA GTC AAC GAT ATT GCA CTA ATC AA - #A GTC AAA TCT CCG ATA          192                                                                       Tyr Asp Ile Val Asn Asp Ile Ala Leu Ile Ly - #s Val Lys Ser Pro Ile                50             - #     55             - #     60                          - - GAA TTC AAT GAA AAA GTA ACG ACT GTA AAA TT - #A GGT GAG GAT TAT GTT          240                                                                       Glu Phe Asn Glu Lys Val Thr Thr Val Lys Le - #u Gly Glu Asp Tyr Val            65                 - # 70                 - # 75                 - # 80       - - GGC GGA GAC GTC CAA CTT CGA TTG ACA GGA TG - #G GGA GTT ACG ACA AAT          288                                                                       Gly Gly Asp Val Gln Leu Arg Leu Thr Gly Tr - #p Gly Val Thr Thr Asn                            85 - #                 90 - #                 95              - - GAG GGA ATC GGA AGC CCG AGT CAA AAA TTA CA - #G GTC ATG ACA GCC AAA          336                                                                       Glu Gly Ile Gly Ser Pro Ser Gln Lys Leu Gl - #n Val Met Thr Ala Lys                       100      - #           105      - #           110                  - - TCA CTA ACT TAT GAG GAT TGC AAA AAC GCA AT - #T TAT AAA AAA GAC TTT          384                                                                       Ser Leu Thr Tyr Glu Asp Cys Lys Asn Ala Il - #e Tyr Lys Lys Asp Phe                   115          - #       120          - #       125                      - - CGA AAG CCA AAT TTG TGC ACA GGC TA    - #                  - #                 410                                                                     Arg Lys Pro Asn Leu Cys Thr Gly                                                   130              - #   135                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:137:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 136 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:                             - - Trp Val Val Thr Ala Ala His Cys Leu Arg Gl - #y Lys Asp His Leu Leu        1               5 - #                 10 - #                 15              - - Asp Lys Leu Phe Ile Ala Val Gly Leu Thr As - #n Leu Gly Glu Gly Gly                   20     - #             25     - #             30                  - - Thr Val Tyr Pro Val Glu Lys Gly Ile Met Hi - #s Glu Glu Tyr Glu His               35         - #         40         - #         45                      - - Tyr Asp Ile Val Asn Asp Ile Ala Leu Ile Ly - #s Val Lys Ser Pro Ile           50             - #     55             - #     60                          - - Glu Phe Asn Glu Lys Val Thr Thr Val Lys Le - #u Gly Glu Asp Tyr Val       65                 - # 70                 - # 75                 - # 80       - - Gly Gly Asp Val Gln Leu Arg Leu Thr Gly Tr - #p Gly Val Thr Thr Asn                       85 - #                 90 - #                 95              - - Glu Gly Ile Gly Ser Pro Ser Gln Lys Leu Gl - #n Val Met Thr Ala Lys                  100      - #           105      - #           110                  - - Ser Leu Thr Tyr Glu Asp Cys Lys Asn Ala Il - #e Tyr Lys Lys Asp Phe              115          - #       120          - #       125                      - - Arg Lys Pro Asn Leu Cys Thr Gly                                              130              - #   135                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:138:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 864 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..781                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 456, change G to                      K; at - #pos. bp 504, change A to R.  At pos. aa - #152       and                                                                                            168, subs - #titute Xaa."                                       - -         (xi) SEQUENCE DESCRIPTION: SEQ - #ID NO:138:                      - - T ACA AAA CCT ATT ACA ATT CAA AAG TTG TTC - #CAA ATG ATG GCA AAT             46                                                                        - -   Thr Lys Pro Ile Thr Ile Gln Lys Leu P - #he Gln Met Met Ala Asn            1              - # 5                 - # 10                 - # 15         - - TTT GTG CTA TTC ACC TTA CTA GCC TTA GTA TC - #A GTA GCA TGT TCC AAA           94                                                                       Phe Val Leu Phe Thr Leu Leu Ala Leu Val Se - #r Val Ala Cys Ser Lys                            20 - #                 25 - #                 30              - - TAT ATT GAT CCA AGA ATC ATT GGA GGC GAA GA - #T GCT CCT GAA GGC TCG          142                                                                       Tyr Ile Asp Pro Arg Ile Ile Gly Gly Glu As - #p Ala Pro Glu Gly Ser                        35     - #             40     - #             45                  - - GCT CCG TAT CAA GTT TCA CTG AGA AAT CGG GA - #C CTA GAG CAT TTC TGT          190                                                                       Ala Pro Tyr Gln Val Ser Leu Arg Asn Arg As - #p Leu Glu His Phe Cys                    50         - #         55         - #         60                      - - GGT GGC TCC ATC TTA AAC AAA CGT TGG ATT GT - #G ACG GCT GCA CAT TGC          238                                                                       Gly Gly Ser Ile Leu Asn Lys Arg Trp Ile Va - #l Thr Ala Ala His Cys                65             - #     70             - #     75                          - - CTA AAA CCT GGC ATT TTA AAA TCC GTC TAT AT - #G GGA TCA AAC TCA TTA          286                                                                       Leu Lys Pro Gly Ile Leu Lys Ser Val Tyr Me - #t Gly Ser Asn Ser Leu            80                 - # 85                 - # 90                 - # 95       - - GAT GGC AAT GGT ACA TAC TAC GAC GTC GAA CG - #T TTT GTG ATG CAT CAT          334                                                                       Asp Gly Asn Gly Thr Tyr Tyr Asp Val Glu Ar - #g Phe Val Met His His                           100  - #               105  - #               110              - - AAA TAT ACA CCA AAA ATT ACT GTC AAC TAT GC - #T GAT ATT GGT CTA ATA          382                                                                       Lys Tyr Thr Pro Lys Ile Thr Val Asn Tyr Al - #a Asp Ile Gly Leu Ile                       115      - #           120      - #           125                  - - AAA GTG ACA AAA GAC ATT ATA TTC AGT GAC AA - #A GTT CAA CCA ATC AAA          430                                                                       Lys Val Thr Lys Asp Ile Ile Phe Ser Asp Ly - #s Val Gln Pro Ile Lys                   130          - #       135          - #       140                      - - ATA GCA AAA AAA ATA TCA AGG GTG GKG AAT CT - #G CAA GGC CAC TGG TTG          478                                                                       Ile Ala Lys Lys Ile Ser Arg Val Xaa Asn Le - #u Gln Gly His Trp Leu               145              - #   150              - #   155                          - - GGG TCG ATT GGC GGA TGG GGC CCC CRG TAC CA - #A ACG AAT TGC AAC AAG          526                                                                       Gly Ser Ile Gly Gly Trp Gly Pro Xaa Tyr Gl - #n Thr Asn Cys Asn Lys           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - GTG GAA ACC ACT GCA ATA ACA AAT GAA AAG TG - #C TAC GAA TTG TCT        CAA      574                                                                    Val Glu Thr Thr Ala Ile Thr Asn Glu Lys Cy - #s Tyr Glu Leu Ser Gln                          180  - #               185  - #               190              - - TTC GTT GAG CCA ACT TCG CAA ATA TGT ACA TT - #A AGG GAA TTT TTA AGA          622                                                                       Phe Val Glu Pro Thr Ser Gln Ile Cys Thr Le - #u Arg Glu Phe Leu Arg                       195      - #           200      - #           205                  - - GGC ATT TGC TTT GGT GAT TCT GGT GGA CCA CT - #G GTT TAC AAA GGT GAA          670                                                                       Gly Ile Cys Phe Gly Asp Ser Gly Gly Pro Le - #u Val Tyr Lys Gly Glu                   210          - #       215          - #       220                      - - CTG GTT GGC GTT TCT TCG TTT GTC TTG TAC AC - #T TGC GGA GCT GGA CGC          718                                                                       Leu Val Gly Val Ser Ser Phe Val Leu Tyr Th - #r Cys Gly Ala Gly Arg               225              - #   230              - #   235                          - - CCA GAT GTT TTT GTT AAA GTG CGT GAT TTC CA - #A TCT TGG ATC AAT TCT          766                                                                       Pro Asp Val Phe Val Lys Val Arg Asp Phe Gl - #n Ser Trp Ile Asn Ser           240                 2 - #45                 2 - #50                 2 -      #55                                                                              - - GAA ATT AGA AAA AAA TAAATAGATT TCAATCATGA TTTGTTGTA - #A TAAAAAATGG          821                                                                      Glu Ile Arg Lys Lys                                                                           260                                                            - - TTAAATAAAG GCAGCATAAT TTAAAAAAAA AAAAAAAAAA AAA    - #                      - #864                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:139:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 260 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:                             - - Thr Lys Pro Ile Thr Ile Gln Lys Leu Phe Gl - #n Met Met Ala Asn Phe        1               5 - #                 10 - #                 15              - - Val Leu Phe Thr Leu Leu Ala Leu Val Ser Va - #l Ala Cys Ser Lys Tyr                   20     - #             25     - #             30                  - - Ile Asp Pro Arg Ile Ile Gly Gly Glu Asp Al - #a Pro Glu Gly Ser Ala               35         - #         40         - #         45                      - - Pro Tyr Gln Val Ser Leu Arg Asn Arg Asp Le - #u Glu His Phe Cys Gly           50             - #     55             - #     60                          - - Gly Ser Ile Leu Asn Lys Arg Trp Ile Val Th - #r Ala Ala His Cys Leu       65                 - # 70                 - # 75                 - # 80       - - Lys Pro Gly Ile Leu Lys Ser Val Tyr Met Gl - #y Ser Asn Ser Leu Asp                       85 - #                 90 - #                 95              - - Gly Asn Gly Thr Tyr Tyr Asp Val Glu Arg Ph - #e Val Met His His Lys                  100      - #           105      - #           110                  - - Tyr Thr Pro Lys Ile Thr Val Asn Tyr Ala As - #p Ile Gly Leu Ile Lys              115          - #       120          - #       125                      - - Val Thr Lys Asp Ile Ile Phe Ser Asp Lys Va - #l Gln Pro Ile Lys Ile          130              - #   135              - #   140                          - - Ala Lys Lys Ile Ser Arg Val Xaa Asn Leu Gl - #n Gly His Trp Leu Gly      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ser Ile Gly Gly Trp Gly Pro Xaa Tyr Gln Th - #r Asn Cys Asn Lys        Val                                                                                             165  - #               170  - #               175             - - Glu Thr Thr Ala Ile Thr Asn Glu Lys Cys Ty - #r Glu Leu Ser Gln Phe                  180      - #           185      - #           190                  - - Val Glu Pro Thr Ser Gln Ile Cys Thr Leu Ar - #g Glu Phe Leu Arg Gly              195          - #       200          - #       205                      - - Ile Cys Phe Gly Asp Ser Gly Gly Pro Leu Va - #l Tyr Lys Gly Glu Leu          210              - #   215              - #   220                          - - Val Gly Val Ser Ser Phe Val Leu Tyr Thr Cy - #s Gly Ala Gly Arg Pro      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asp Val Phe Val Lys Val Arg Asp Phe Gln Se - #r Trp Ile Asn Ser        Glu                                                                                             245  - #               250  - #               255             - - Ile Arg Lys Lys                                                                      260                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:140:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 610 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..555                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 133, change A to                      H; at - #pos. bp 168, change A to W.  At pos. aa - #45,                       substitute - #Xaa."                                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140:                             - - GTT TTG ACA GCT GCT CAT TGT GTA GAA AAG TA - #T AAA TTG ACT GTT AGA           48                                                                       Val Leu Thr Ala Ala His Cys Val Glu Lys Ty - #r Lys Leu Thr Val Arg             1               5 - #                 10 - #                 15              - - GTT GGC AGC AGC GAT TTG GAA TCA GGA GGC AA - #A ATA CAT ACC ATT AAA           96                                                                       Val Gly Ser Ser Asp Leu Glu Ser Gly Gly Ly - #s Ile His Thr Ile Lys                        20     - #             25     - #             30                  - - AAA ATC CAT GTT CAT CCA TAT TAC GAA CCG GT - #T GAC HAC GAT TTT GCC          144                                                                       Lys Ile His Val His Pro Tyr Tyr Glu Pro Va - #l Asp Xaa Asp Phe Ala                    35         - #         40         - #         45                      - - TTG CTG GGA CTT GAT GAA CCA GTW TTC TTG AG - #C AAC AAA GTT CAA CTT          192                                                                       Leu Leu Gly Leu Asp Glu Pro Val Phe Leu Se - #r Asn Lys Val Gln Leu                50             - #     55             - #     60                          - - GTG AAA CTT GTA GAA CAA GGT GTA GAT CTG GA - #T GAA GGA ACC TTT CTT          240                                                                       Val Lys Leu Val Glu Gln Gly Val Asp Leu As - #p Glu Gly Thr Phe Leu            65                 - # 70                 - # 75                 - # 80       - - AAT GCC ACT GGA TGG GGT ACA ACA GCG ACC GA - #A GAT TTG GCT CCA GTT          288                                                                       Asn Ala Thr Gly Trp Gly Thr Thr Ala Thr Gl - #u Asp Leu Ala Pro Val                            85 - #                 90 - #                 95              - - CTT CAA TTA GTA ACA GTT CCA GTA GTC AAC AC - #A TAT ACT TGC AGC AAA          336                                                                       Leu Gln Leu Val Thr Val Pro Val Val Asn Th - #r Tyr Thr Cys Ser Lys                       100      - #           105      - #           110                  - - ATT TAC GAC TTT GGT ATC ACA CAA AGA ATG TT - #T TGC GCT GGT TAT ATG          384                                                                       Ile Tyr Asp Phe Gly Ile Thr Gln Arg Met Ph - #e Cys Ala Gly Tyr Met                   115          - #       120          - #       125                      - - GAT GGA ACT CTT AAG GAC ATC TGC TCT GGA GA - #T TCA GGT AGT CCT GTG          432                                                                       Asp Gly Thr Leu Lys Asp Ile Cys Ser Gly As - #p Ser Gly Ser Pro Val               130              - #   135              - #   140                          - - GTG AAG GAT GGT ATC CAA TAT GGT GTG GTG TC - #T TGG GGA AAA GCT TGT          480                                                                       Val Lys Asp Gly Ile Gln Tyr Gly Val Val Se - #r Trp Gly Lys Ala Cys           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - GCC GAT CCA AGA TAT CCA AAT GTT TAT TCC AA - #A GTT AGC TAC GAA        CGT      528                                                                    Ala Asp Pro Arg Tyr Pro Asn Val Tyr Ser Ly - #s Val Ser Tyr Glu Arg                          165  - #               170  - #               175              - - ATA TGG ATT AAA GAA GTG TCT GGA GTT TAAGTAGAT - #G CATTCTTTAT                575                                                                       Ile Trp Ile Lys Glu Val Ser Gly Val                                                       180      - #           185                                         - - TATAATAAAA TTGTTTAATT AAAAAAAAAA AAAAA       - #                  -     #      610                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:141:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 185 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:                             - - Val Leu Thr Ala Ala His Cys Val Glu Lys Ty - #r Lys Leu Thr Val Arg        1               5 - #                 10 - #                 15              - - Val Gly Ser Ser Asp Leu Glu Ser Gly Gly Ly - #s Ile His Thr Ile Lys                   20     - #             25     - #             30                  - - Lys Ile His Val His Pro Tyr Tyr Glu Pro Va - #l Asp Xaa Asp Phe Ala               35         - #         40         - #         45                      - - Leu Leu Gly Leu Asp Glu Pro Val Phe Leu Se - #r Asn Lys Val Gln Leu           50             - #     55             - #     60                          - - Val Lys Leu Val Glu Gln Gly Val Asp Leu As - #p Glu Gly Thr Phe Leu       65                 - # 70                 - # 75                 - # 80       - - Asn Ala Thr Gly Trp Gly Thr Thr Ala Thr Gl - #u Asp Leu Ala Pro Val                       85 - #                 90 - #                 95              - - Leu Gln Leu Val Thr Val Pro Val Val Asn Th - #r Tyr Thr Cys Ser Lys                  100      - #           105      - #           110                  - - Ile Tyr Asp Phe Gly Ile Thr Gln Arg Met Ph - #e Cys Ala Gly Tyr Met              115          - #       120          - #       125                      - - Asp Gly Thr Leu Lys Asp Ile Cys Ser Gly As - #p Ser Gly Ser Pro Val          130              - #   135              - #   140                          - - Val Lys Asp Gly Ile Gln Tyr Gly Val Val Se - #r Trp Gly Lys Ala Cys      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ala Asp Pro Arg Tyr Pro Asn Val Tyr Ser Ly - #s Val Ser Tyr Glu        Arg                                                                                             165  - #               170  - #               175             - - Ile Trp Ile Lys Glu Val Ser Gly Val                                                  180      - #           185                                         - -  - - (2) INFORMATION FOR SEQ ID NO:142:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 386 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..384                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 3, change A to                        R."                                                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142:                             - - AAR ATT GGT ACT AGC CAT AGG ATA CTT TAT GG - #T AGA GTG ATA GAC ATA           48                                                                       Lys Ile Gly Thr Ser His Arg Ile Leu Tyr Gl - #y Arg Val Ile Asp Ile             1               5 - #                 10 - #                 15              - - AAA GAA ATT ATA ATG CAT CCA GAC TAT ACG TC - #T GTA TCA GGA AGT GGA           96                                                                       Lys Glu Ile Ile Met His Pro Asp Tyr Thr Se - #r Val Ser Gly Ser Gly                        20     - #             25     - #             30                  - - TAC GAT GTA GCA CTA TTG AAA CCG TCT ACA AA - #A ATT GTT TTT AAC TCA          144                                                                       Tyr Asp Val Ala Leu Leu Lys Pro Ser Thr Ly - #s Ile Val Phe Asn Ser                    35         - #         40         - #         45                      - - AAA TCT ATC AAA CCT GTA AAG CTA ATT GAT GA - #A GGA ATC GAA ACG GCT          192                                                                       Lys Ser Ile Lys Pro Val Lys Leu Ile Asp Gl - #u Gly Ile Glu Thr Ala                50             - #     55             - #     60                          - - AAT GGT TCA ATA GCA ACC GTG GCA GGT TGG GG - #T AAA GTA GTG GAT GGT          240                                                                       Asn Gly Ser Ile Ala Thr Val Ala Gly Trp Gl - #y Lys Val Val Asp Gly            65                 - # 70                 - # 75                 - # 80       - - TTT CCG TAC ATA CCT AAT TAT TTA TTG GCT GT - #A AAT GTA CCG ATT ATT          288                                                                       Phe Pro Tyr Ile Pro Asn Tyr Leu Leu Ala Va - #l Asn Val Pro Ile Ile                            85 - #                 90 - #                 95              - - GAT AGC GAT ACA TGC AAG TCG ATG AAT ATT GA - #A TAT CAA AAA TAT TTG          336                                                                       Asp Ser Asp Thr Cys Lys Ser Met Asn Ile Gl - #u Tyr Gln Lys Tyr Leu                       100      - #           105      - #           110                  - - AAA CCA AAT ATG ATA TGC GCC GGA TAT GCA AA - #A GGT GGT AAA GAT TCT          384                                                                       Lys Pro Asn Met Ile Cys Ala Gly Tyr Ala Ly - #s Gly Gly Lys Asp Ser                   115          - #       120          - #       125                      - - TG                  - #                  - #                  - #                 386                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:143:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 128 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:                             - - Lys Ile Gly Thr Ser His Arg Ile Leu Tyr Gl - #y Arg Val Ile Asp Ile        1               5 - #                 10 - #                 15              - - Lys Glu Ile Ile Met His Pro Asp Tyr Thr Se - #r Val Ser Gly Ser Gly                   20     - #             25     - #             30                  - - Tyr Asp Val Ala Leu Leu Lys Pro Ser Thr Ly - #s Ile Val Phe Asn Ser               35         - #         40         - #         45                      - - Lys Ser Ile Lys Pro Val Lys Leu Ile Asp Gl - #u Gly Ile Glu Thr Ala           50             - #     55             - #     60                          - - Asn Gly Ser Ile Ala Thr Val Ala Gly Trp Gl - #y Lys Val Val Asp Gly       65                 - # 70                 - # 75                 - # 80       - - Phe Pro Tyr Ile Pro Asn Tyr Leu Leu Ala Va - #l Asn Val Pro Ile Ile                       85 - #                 90 - #                 95              - - Asp Ser Asp Thr Cys Lys Ser Met Asn Ile Gl - #u Tyr Gln Lys Tyr Leu                  100      - #           105      - #           110                  - - Lys Pro Asn Met Ile Cys Ala Gly Tyr Ala Ly - #s Gly Gly Lys Asp Ser              115          - #       120          - #       125                      - -  - - (2) INFORMATION FOR SEQ ID NO:144:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 923 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..802                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 896, change G to                      B; at - #pos. bp 899, change A to W."                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144:                             - - A TTA ACA ATG ATG AAA CTT TTG GTA GTT TTT - #GCG ATT TTC GCT CAA             46                                                                          Leu Thr Met Met Lys Leu Leu Val Val P - #he Ala Ile Phe Ala Gln                 1              - # 5                 - # 10                 - # 15         - - ATC AGT TTT GTT TTT GGA AAT AAT GTA ACA GA - #A TTC GAT GAC CGA ATC           94                                                                       Ile Ser Phe Val Phe Gly Asn Asn Val Thr Gl - #u Phe Asp Asp Arg Ile                            20 - #                 25 - #                 30              - - GTT GGA GGT GAA GAT GTT GAT ATA TCA ACT TG - #T GGT TGG CAA ATT TCA          142                                                                       Val Gly Gly Glu Asp Val Asp Ile Ser Thr Cy - #s Gly Trp Gln Ile Ser                        35     - #             40     - #             45                  - - TTT CAA AGT GAA AAC CTT CAT TTT TGT GGA GG - #A TCA ATT ATT GCA CCA          190                                                                       Phe Gln Ser Glu Asn Leu His Phe Cys Gly Gl - #y Ser Ile Ile Ala Pro                    50         - #         55         - #         60                      - - AAA TGG ATT CTA ACT GCT GCA CAC TGT GTT GA - #A TGG TTG AAA AAG CCG          238                                                                       Lys Trp Ile Leu Thr Ala Ala His Cys Val Gl - #u Trp Leu Lys Lys Pro                65             - #     70             - #     75                          - - CTC AAA GAC ATA ACC GTA CGT ATA GGA AGC AG - #T ATA CGT AAC AAA GGT          286                                                                       Leu Lys Asp Ile Thr Val Arg Ile Gly Ser Se - #r Ile Arg Asn Lys Gly            80                 - # 85                 - # 90                 - # 95       - - GGT CGA GTT CAT AAA GTA ATA GAT TTC CAC AT - #G CAT CCC TCG TAC AAT          334                                                                       Gly Arg Val His Lys Val Ile Asp Phe His Me - #t His Pro Ser Tyr Asn                           100  - #               105  - #               110              - - AAG AGG GCG GAT TAT GAT TTT GAC GTT GCT GT - #A CTA GAA CTT GAA AAA          382                                                                       Lys Arg Ala Asp Tyr Asp Phe Asp Val Ala Va - #l Leu Glu Leu Glu Lys                       115      - #           120      - #           125                  - - CCA GTC TCA TAT ACG GTT TGT ACA GTA GTA TC - #A GTA GAT TTA GCC GAA          430                                                                       Pro Val Ser Tyr Thr Val Cys Thr Val Val Se - #r Val Asp Leu Ala Glu                   130          - #       135          - #       140                      - - AGT GGA ACT GAA GTT AAA CCT GGA GCA ATA CT - #T AGT GTC ACT GGA TGG          478                                                                       Ser Gly Thr Glu Val Lys Pro Gly Ala Ile Le - #u Ser Val Thr Gly Trp               145              - #   150              - #   155                          - - GGT GCA ACT AAG GAA GGT GGT GGC GGA ACT TT - #G CAA CTA CAA GGT GTG          526                                                                       Gly Ala Thr Lys Glu Gly Gly Gly Gly Thr Le - #u Gln Leu Gln Gly Val           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - AAA GTT CCA GCT ATC TCT CCC AAA GAT TGT GC - #T AAG GGA TAT CCA        CCT      574                                                                    Lys Val Pro Ala Ile Ser Pro Lys Asp Cys Al - #a Lys Gly Tyr Pro Pro                          180  - #               185  - #               190              - - TCT GGA GGT AAA GAC AAA ATT ACA GAC AGC AT - #G TTA TGT GCT GGT CTT          622                                                                       Ser Gly Gly Lys Asp Lys Ile Thr Asp Ser Me - #t Leu Cys Ala Gly Leu                       195      - #           200      - #           205                  - - CCT GAA GGA GGT AAA GAT TCC TGC CAA GGC GA - #C AGT GGC GGT CCA CTG          670                                                                       Pro Glu Gly Gly Lys Asp Ser Cys Gln Gly As - #p Ser Gly Gly Pro Leu                   210          - #       215          - #       220                      - - GTA GAT GAA AAT AGA AAG CAA GTA GGA GTG GT - #T TCT TGG GGT CAA GGA          718                                                                       Val Asp Glu Asn Arg Lys Gln Val Gly Val Va - #l Ser Trp Gly Gln Gly               225              - #   230              - #   235                          - - TGT GCC AGA CCA GGA AAA CCA GGA ATT TAT GC - #T AAA GTG TCA CAC CCC          766                                                                       Cys Ala Arg Pro Gly Lys Pro Gly Ile Tyr Al - #a Lys Val Ser His Pro           240                 2 - #45                 2 - #50                 2 -      #55                                                                              - - GAA ATC AGA AAA TTT ATT GAA AAA TAT GCT AA - #T GTT TAAGTGGATT               812                                                                      Glu Ile Arg Lys Phe Ile Glu Lys Tyr Ala As - #n Val                                           260  - #               265                                     - - TCATTTTCAA TATAATGTGA TTTAAGATAC TCTTTAATGT TATGATATGA AT -             #TGTGATAA    872                                                                 - - ATTAAATAAT AAAGATTGAA GAABTGWTAA AAAAAAAAAA AAAAAAAAAA A - #                923                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:145:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 267 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:                             - - Leu Thr Met Met Lys Leu Leu Val Val Phe Al - #a Ile Phe Ala Gln Ile        1               5 - #                 10 - #                 15              - - Ser Phe Val Phe Gly Asn Asn Val Thr Glu Ph - #e Asp Asp Arg Ile Val                   20     - #             25     - #             30                  - - Gly Gly Glu Asp Val Asp Ile Ser Thr Cys Gl - #y Trp Gln Ile Ser Phe               35         - #         40         - #         45                      - - Gln Ser Glu Asn Leu His Phe Cys Gly Gly Se - #r Ile Ile Ala Pro Lys           50             - #     55             - #     60                          - - Trp Ile Leu Thr Ala Ala His Cys Val Glu Tr - #p Leu Lys Lys Pro Leu       65                 - # 70                 - # 75                 - # 80       - - Lys Asp Ile Thr Val Arg Ile Gly Ser Ser Il - #e Arg Asn Lys Gly Gly                       85 - #                 90 - #                 95              - - Arg Val His Lys Val Ile Asp Phe His Met Hi - #s Pro Ser Tyr Asn Lys                  100      - #           105      - #           110                  - - Arg Ala Asp Tyr Asp Phe Asp Val Ala Val Le - #u Glu Leu Glu Lys Pro              115          - #       120          - #       125                      - - Val Ser Tyr Thr Val Cys Thr Val Val Ser Va - #l Asp Leu Ala Glu Ser          130              - #   135              - #   140                          - - Gly Thr Glu Val Lys Pro Gly Ala Ile Leu Se - #r Val Thr Gly Trp Gly      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ala Thr Lys Glu Gly Gly Gly Gly Thr Leu Gl - #n Leu Gln Gly Val        Lys                                                                                             165  - #               170  - #               175             - - Val Pro Ala Ile Ser Pro Lys Asp Cys Ala Ly - #s Gly Tyr Pro Pro Ser                  180      - #           185      - #           190                  - - Gly Gly Lys Asp Lys Ile Thr Asp Ser Met Le - #u Cys Ala Gly Leu Pro              195          - #       200          - #       205                      - - Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp Se - #r Gly Gly Pro Leu Val          210              - #   215              - #   220                          - - Asp Glu Asn Arg Lys Gln Val Gly Val Val Se - #r Trp Gly Gln Gly Cys      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Ala Arg Pro Gly Lys Pro Gly Ile Tyr Ala Ly - #s Val Ser His Pro        Glu                                                                                             245  - #               250  - #               255             - - Ile Arg Lys Phe Ile Glu Lys Tyr Ala Asn Va - #l                                      260      - #           265                                         - -  - - (2) INFORMATION FOR SEQ ID NO:146:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 612 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..612                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:                             - - GTT TGC AAG ATG GCA ACA ACT CTG TTT TCT CT - #T GCC ATA GTG TTG TTT           48                                                                       Val Cys Lys Met Ala Thr Thr Leu Phe Ser Le - #u Ala Ile Val Leu Phe             1               5 - #                 10 - #                 15              - - ATC TCT ACT TCA GAA GAA TCA GCA CAT ATT TC - #G CAA GGA TCT CGA ATA           96                                                                       Ile Ser Thr Ser Glu Glu Ser Ala His Ile Se - #r Gln Gly Ser Arg Ile                        20     - #             25     - #             30                  - - TTA GGA GGT AGA AAT GCA AAA CTC GGA GAT GC - #T CCT TAT CAA GTA TCA          144                                                                       Leu Gly Gly Arg Asn Ala Lys Leu Gly Asp Al - #a Pro Tyr Gln Val Ser                    35         - #         40         - #         45                      - - CTA AGA GAT AAT TTT GGA CAT TTT TGT GGA GG - #T TCT ATC ATT AGT GAA          192                                                                       Leu Arg Asp Asn Phe Gly His Phe Cys Gly Gl - #y Ser Ile Ile Ser Glu                50             - #     55             - #     60                          - - AAT TTT GTG ATT ACA GCA GCT CAT TGC CTT GA - #T GGA TAC ACA GTG AGC          240                                                                       Asn Phe Val Ile Thr Ala Ala His Cys Leu As - #p Gly Tyr Thr Val Ser            65                 - # 70                 - # 75                 - # 80       - - AAA TTT AAA GTA GCA ACT GGT ACA ATC GAG TA - #T GGT AAA GGA GGT GAC          288                                                                       Lys Phe Lys Val Ala Thr Gly Thr Ile Glu Ty - #r Gly Lys Gly Gly Asp                            85 - #                 90 - #                 95              - - GAA TAT AAA GTC ATC AAC TTT GTT GTC CGG GA - #T GAC TTT CAA TAT GTC          336                                                                       Glu Tyr Lys Val Ile Asn Phe Val Val Arg As - #p Asp Phe Gln Tyr Val                       100      - #           105      - #           110                  - - AAA TTA GAA AAT GAT ATT GCT ATA GTA CAA AT - #A GAT GGA TCT TTT AAA          384                                                                       Lys Leu Glu Asn Asp Ile Ala Ile Val Gln Il - #e Asp Gly Ser Phe Lys                   115          - #       120          - #       125                      - - TTT AAT GAC TAT GTA AAG CCT ATA AAA TTG CC - #A AAT CAA GAT ACT AAA          432                                                                       Phe Asn Asp Tyr Val Lys Pro Ile Lys Leu Pr - #o Asn Gln Asp Thr Lys               130              - #   135              - #   140                          - - GTT GGC GCG GAT GTT GTC CTA ACA GGA TGG GG - #A AAA ATG GAA GGT GGT          480                                                                       Val Gly Ala Asp Val Val Leu Thr Gly Trp Gl - #y Lys Met Glu Gly Gly           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - AAA AAT CCA GAA ACT CTA CAA ATC TTG AAC TT - #A AAA ACA ATT GAT        CAA      528                                                                    Lys Asn Pro Glu Thr Leu Gln Ile Leu Asn Le - #u Lys Thr Ile Asp Gln                          165  - #               170  - #               175              - - GGA GAA TGC AAA CAA GCT TTG GCA GAA GTA AA - #C ACA GTT CTT CCA AGT          576                                                                       Gly Glu Cys Lys Gln Ala Leu Ala Glu Val As - #n Thr Val Leu Pro Ser                       180      - #           185      - #           190                  - - CAA ATT TGT ACC TAT GTT GGT GTT GGC AAA GG - #A GCT                     - #      612                                                                    Gln Ile Cys Thr Tyr Val Gly Val Gly Lys Gl - #y Ala                                   195          - #       200                                             - -  - - (2) INFORMATION FOR SEQ ID NO:147:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 204 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:                             - - Val Cys Lys Met Ala Thr Thr Leu Phe Ser Le - #u Ala Ile Val Leu        Phe                                                                               1               5 - #                 10 - #                 15             - - Ile Ser Thr Ser Glu Glu Ser Ala His Ile Se - #r Gln Gly Ser Arg Ile                   20     - #             25     - #             30                  - - Leu Gly Gly Arg Asn Ala Lys Leu Gly Asp Al - #a Pro Tyr Gln Val Ser               35         - #         40         - #         45                      - - Leu Arg Asp Asn Phe Gly His Phe Cys Gly Gl - #y Ser Ile Ile Ser Glu           50             - #     55             - #     60                          - - Asn Phe Val Ile Thr Ala Ala His Cys Leu As - #p Gly Tyr Thr Val Ser       65                 - # 70                 - # 75                 - # 80       - - Lys Phe Lys Val Ala Thr Gly Thr Ile Glu Ty - #r Gly Lys Gly Gly Asp                       85 - #                 90 - #                 95              - - Glu Tyr Lys Val Ile Asn Phe Val Val Arg As - #p Asp Phe Gln Tyr Val                  100      - #           105      - #           110                  - - Lys Leu Glu Asn Asp Ile Ala Ile Val Gln Il - #e Asp Gly Ser Phe Lys              115          - #       120          - #       125                      - - Phe Asn Asp Tyr Val Lys Pro Ile Lys Leu Pr - #o Asn Gln Asp Thr Lys          130              - #   135              - #   140                          - - Val Gly Ala Asp Val Val Leu Thr Gly Trp Gl - #y Lys Met Glu Gly Gly      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Lys Asn Pro Glu Thr Leu Gln Ile Leu Asn Le - #u Lys Thr Ile Asp        Gln                                                                                             165  - #               170  - #               175             - - Gly Glu Cys Lys Gln Ala Leu Ala Glu Val As - #n Thr Val Leu Pro Ser                  180      - #           185      - #           190                  - - Gln Ile Cys Thr Tyr Val Gly Val Gly Lys Gl - #y Ala                              195          - #       200                                             - -  - - (2) INFORMATION FOR SEQ ID NO:148:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 641 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..641                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 91, change G to                       N; at - #pos. bp 385, change A to M; at bp 404, ch -          #ange                                                                                          A to W - #. At pos. aa 30, 128 and 134, substitute Xaa -     #."                                                                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:                             - - CA AAC ATG AAA CTT TAT GTT TTT ACA TTG ATC - # ATC GCA GCC GCT GTT            47                                                                          Asn Met Lys Leu Tyr Val Phe Thr Leu - #Ile Ile Ala Ala Ala Val                  1             - #  5                - #  10                - #  15        - - GCT GCA CCA ACA AAC GAA ATC GAT TGG ATA GG - #C TTT CCC GAA GNT TTT           95                                                                       Ala Ala Pro Thr Asn Glu Ile Asp Trp Ile Gl - #y Phe Pro Glu Xaa Phe                            20 - #                 25 - #                 30              - - CCT CGT GTA GTT GGT GGC CAG ACT GCC AAA CC - #T CAC CAG TTC CCC TGG          143                                                                       Pro Arg Val Val Gly Gly Gln Thr Ala Lys Pr - #o His Gln Phe Pro Trp                        35     - #             40     - #             45                  - - CAG GTT TCC CTG CAA AGG TCC GGA AAG CAT TT - #G TGC GGC GGT TCC ATC          191                                                                       Gln Val Ser Leu Gln Arg Ser Gly Lys His Le - #u Cys Gly Gly Ser Ile                    50         - #         55         - #         60                      - - TTG AAC GAC AGG TGG GTC TTG ACA GCC GCG CA - #C TGC ATC AGC GGA ACT          239                                                                       Leu Asn Asp Arg Trp Val Leu Thr Ala Ala Hi - #s Cys Ile Ser Gly Thr                65             - #     70             - #     75                          - - GAA AAT TAC GAG GCT GTA GTC GGA AAA CAC GA - #T TTG TCG AAA AGC GAA          287                                                                       Glu Asn Tyr Glu Ala Val Val Gly Lys His As - #p Leu Ser Lys Ser Glu            80                 - # 85                 - # 90                 - # 95       - - TCA TCT GAG CAA CGT TGC GCC TAC AAG AGG AC - #C ATC GTC CAC TCG TCC          335                                                                       Ser Ser Glu Gln Arg Cys Ala Tyr Lys Arg Th - #r Ile Val His Ser Ser                           100  - #               105  - #               110              - - TTC ACT GGA AGG GTG GGT CCT TAC GAT GTC GC - #T TTG ATT GAA TTA GAA          383                                                                       Phe Thr Gly Arg Val Gly Pro Tyr Asp Val Al - #a Leu Ile Glu Leu Glu                       115      - #           120      - #           125                  - - AMA CCT TTC AAA TTG AAC GAW AAA TGT AAG CC - #A ATC AGA CTT CCA TTG          431                                                                       Xaa Pro Phe Lys Leu Asn Xaa Lys Cys Lys Pr - #o Ile Arg Leu Pro Leu                   130          - #       135          - #       140                      - - AAA GAT GAA GCA CAT TCT GGA CAA GTG ACG CT - #T TCT GGT TGG GGA TCT          479                                                                       Lys Asp Glu Ala His Ser Gly Gln Val Thr Le - #u Ser Gly Trp Gly Ser               145              - #   150              - #   155                          - - ACT TCA ACT ACC ATC TTC CCC ACT TAC CCA AA - #T GAA CTA CAG TAT GTT          527                                                                       Thr Ser Thr Thr Ile Phe Pro Thr Tyr Pro As - #n Glu Leu Gln Tyr Val           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - GAC AAA CCG ATT GTT CCA TAT ACT GAT TGC GA - #A AAT GCT ATG GGC        GGA      575                                                                    Asp Lys Pro Ile Val Pro Tyr Thr Asp Cys Gl - #u Asn Ala Met Gly Gly                          180  - #               185  - #               190              - - CCA GGA GCA TCT CCT CTT GAT CCT TTG AAC AT - #C TGC ACT GGT CCC TTG          623                                                                       Pro Gly Ala Ser Pro Leu Asp Pro Leu Asn Il - #e Cys Thr Gly Pro Leu                       195      - #           200      - #           205                  - - ACT GGT GGC ATC AGT GCT         - #                  - #                      - # 641                                                                  Thr Gly Gly Ile Ser Ala                                                               210                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:149:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 213 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:                             - - Asn Met Lys Leu Tyr Val Phe Thr Leu Ile Il - #e Ala Ala Ala Val Ala        1               5 - #                 10 - #                 15              - - Ala Pro Thr Asn Glu Ile Asp Trp Ile Gly Ph - #e Pro Glu Xaa Phe Pro                   20     - #             25     - #             30                  - - Arg Val Val Gly Gly Gln Thr Ala Lys Pro Hi - #s Gln Phe Pro Trp Gln               35         - #         40         - #         45                      - - Val Ser Leu Gln Arg Ser Gly Lys His Leu Cy - #s Gly Gly Ser Ile Leu           50             - #     55             - #     60                          - - Asn Asp Arg Trp Val Leu Thr Ala Ala His Cy - #s Ile Ser Gly Thr Glu       65                 - # 70                 - # 75                 - # 80       - - Asn Tyr Glu Ala Val Val Gly Lys His Asp Le - #u Ser Lys Ser Glu Ser                       85 - #                 90 - #                 95              - - Ser Glu Gln Arg Cys Ala Tyr Lys Arg Thr Il - #e Val His Ser Ser Phe                  100      - #           105      - #           110                  - - Thr Gly Arg Val Gly Pro Tyr Asp Val Ala Le - #u Ile Glu Leu Glu Xaa              115          - #       120          - #       125                      - - Pro Phe Lys Leu Asn Xaa Lys Cys Lys Pro Il - #e Arg Leu Pro Leu Lys          130              - #   135              - #   140                          - - Asp Glu Ala His Ser Gly Gln Val Thr Leu Se - #r Gly Trp Gly Ser Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ser Thr Thr Ile Phe Pro Thr Tyr Pro Asn Gl - #u Leu Gln Tyr Val        Asp                                                                                             165  - #               170  - #               175             - - Lys Pro Ile Val Pro Tyr Thr Asp Cys Glu As - #n Ala Met Gly Gly Pro                  180      - #           185      - #           190                  - - Gly Ala Ser Pro Leu Asp Pro Leu Asn Ile Cy - #s Thr Gly Pro Leu Thr              195          - #       200          - #       205                      - - Gly Gly Ile Ser Ala                                                          210                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:150:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 626 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..626                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:                             - - TG ACT ATG AAC TTT GCA TTG TGG TTT GTG ACT - # CTT GTG TCC ATC ACC            47                                                                        - -    Thr Met Asn Phe Ala Leu Trp Phe Val - #Thr Leu Val Ser Ile Thr             1             - #  5                - #  10                - #  15        - - AGC GCT GAT CCA ATT CGT GTT TCC TCA ATT GG - #A AAT GCT AAT ATG GAA           95                                                                       Ser Ala Asp Pro Ile Arg Val Ser Ser Ile Gl - #y Asn Ala Asn Met Glu                            20 - #                 25 - #                 30              - - TCT CGC GTT GTT GGT GGC GAA AAC GCA GAA GT - #A GGA GCT GCT CCT TAC          143                                                                       Ser Arg Val Val Gly Gly Glu Asn Ala Glu Va - #l Gly Ala Ala Pro Tyr                        35     - #             40     - #             45                  - - CAA GTT TCT TTG AAA TAC AAT AAT GGA GCT CA - #T TTT TGC GGA GGT GTC          191                                                                       Gln Val Ser Leu Lys Tyr Asn Asn Gly Ala Hi - #s Phe Cys Gly Gly Val                    50         - #         55         - #         60                      - - GTG ATA ACC AAA ACT TGG GTG CTA ACT GCT GC - #A CGT TGT ATT CAC GAG          239                                                                       Val Ile Thr Lys Thr Trp Val Leu Thr Ala Al - #a Arg Cys Ile His Glu                65             - #     70             - #     75                          - - GAG GAA CCC GAC AGA TTC ACA GTG GTT GTT GG - #T ACC AAC ACT TTA AAT          287                                                                       Glu Glu Pro Asp Arg Phe Thr Val Val Val Gl - #y Thr Asn Thr Leu Asn            80                 - # 85                 - # 90                 - # 95       - - GCT GGA GGA GAA GGT TAC AAC GTT AAA CAG AT - #A GTT ATT CAT ATG CAA          335                                                                       Ala Gly Gly Glu Gly Tyr Asn Val Lys Gln Il - #e Val Ile His Met Gln                           100  - #               105  - #               110              - - TTC AAC CAA GTT TAT CTT CTG AAC GAT ATT GG - #T TTG ATC GAA ACC GAA          383                                                                       Phe Asn Gln Val Tyr Leu Leu Asn Asp Ile Gl - #y Leu Ile Glu Thr Glu                       115      - #           120      - #           125                  - - TCA CCA ATA CAA TTT CAT GAT CTG GTT AAG CC - #A ATC TCA GTC CCC AAT          431                                                                       Ser Pro Ile Gln Phe His Asp Leu Val Lys Pr - #o Ile Ser Val Pro Asn                   130          - #       135          - #       140                      - - ATG CAT GTT GAA GAT GGC ACG AGA GTT ACT CT - #T TTT GGA TGG GGA AAT          479                                                                       Met His Val Glu Asp Gly Thr Arg Val Thr Le - #u Phe Gly Trp Gly Asn               145              - #   150              - #   155                          - - TTA ACG GCT GAA GGA CAT ATG CCA AAC CAT TT - #A CAA ACA ATT GAT TTG          527                                                                       Leu Thr Ala Glu Gly His Met Pro Asn His Le - #u Gln Thr Ile Asp Leu           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - TTA ACA ATA AAT CTG AGT GAA TGC TCC CGA TT - #A TTA CCT GAA CCA        AGT      575                                                                    Leu Thr Ile Asn Leu Ser Glu Cys Ser Arg Le - #u Leu Pro Glu Pro Ser                          180  - #               185  - #               190              - - ATG ATC AGT ACA AAA CAC ATT TGC ACC TTT GT - #T TCA TAT GGA AAA GGA          623                                                                       Met Ile Ser Thr Lys His Ile Cys Thr Phe Va - #l Ser Tyr Gly Lys Gly                       195      - #           200      - #           205                  - - CTT                  - #                  - #                  - #                626                                                                  Leu                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:151:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 208 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:                             - - Thr Met Asn Phe Ala Leu Trp Phe Val Thr Le - #u Val Ser Ile Thr Ser        1               5 - #                 10 - #                 15              - - Ala Asp Pro Ile Arg Val Ser Ser Ile Gly As - #n Ala Asn Met Glu Ser                   20     - #             25     - #             30                  - - Arg Val Val Gly Gly Glu Asn Ala Glu Val Gl - #y Ala Ala Pro Tyr Gln               35         - #         40         - #         45                      - - Val Ser Leu Lys Tyr Asn Asn Gly Ala His Ph - #e Cys Gly Gly Val Val           50             - #     55             - #     60                          - - Ile Thr Lys Thr Trp Val Leu Thr Ala Ala Ar - #g Cys Ile His Glu Glu       65                 - # 70                 - # 75                 - # 80       - - Glu Pro Asp Arg Phe Thr Val Val Val Gly Th - #r Asn Thr Leu Asn Ala                       85 - #                 90 - #                 95              - - Gly Gly Glu Gly Tyr Asn Val Lys Gln Ile Va - #l Ile His Met Gln Phe                  100      - #           105      - #           110                  - - Asn Gln Val Tyr Leu Leu Asn Asp Ile Gly Le - #u Ile Glu Thr Glu Ser              115          - #       120          - #       125                      - - Pro Ile Gln Phe His Asp Leu Val Lys Pro Il - #e Ser Val Pro Asn Met          130              - #   135              - #   140                          - - His Val Glu Asp Gly Thr Arg Val Thr Leu Ph - #e Gly Trp Gly Asn Leu      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Thr Ala Glu Gly His Met Pro Asn His Leu Gl - #n Thr Ile Asp Leu        Leu                                                                                             165  - #               170  - #               175             - - Thr Ile Asn Leu Ser Glu Cys Ser Arg Leu Le - #u Pro Glu Pro Ser Met                  180      - #           185      - #           190                  - - Ile Ser Thr Lys His Ile Cys Thr Phe Val Se - #r Tyr Gly Lys Gly Leu              195          - #       200          - #       205                      - -  - - (2) INFORMATION FOR SEQ ID NO:152:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 433 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..432                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:                             - - GTA ACT GCT GCA CAT TGC TTT TAT GGA ACG TT - #A TTT CCG ATT GGA TTC           48                                                                       Val Thr Ala Ala His Cys Phe Tyr Gly Thr Le - #u Phe Pro Ile Gly Phe             1               5 - #                 10 - #                 15              - - TCT GCG AGA GCC GGC AGC AGT ACT GTG AAT TC - #A GGA GGA ACT GTG CAT           96                                                                       Ser Ala Arg Ala Gly Ser Ser Thr Val Asn Se - #r Gly Gly Thr Val His                        20     - #             25     - #             30                  - - ACA ATT TTG TAT TGG TAT ATT CAT CCA AAT TA - #T GAT TCA CAA AGT ACA          144                                                                       Thr Ile Leu Tyr Trp Tyr Ile His Pro Asn Ty - #r Asp Ser Gln Ser Thr                    35         - #         40         - #         45                      - - GAC TTT GAT GTT TCT GTA GTT CGA CTA TTA TC - #T TCT TTA AAT TTG AAT          192                                                                       Asp Phe Asp Val Ser Val Val Arg Leu Leu Se - #r Ser Leu Asn Leu Asn                50             - #     55             - #     60                          - - GGA GGT TCT ATT CGA CCG GCT AGG TTA GTG GA - #T TCT GGA ACT GAT TTG          240                                                                       Gly Gly Ser Ile Arg Pro Ala Arg Leu Val As - #p Ser Gly Thr Asp Leu            65                 - # 70                 - # 75                 - # 80       - - CCA GCC GGT GAG ATG GTT ACA GTA ACT GGA TG - #G GGA CGA CTT TCG GAA          288                                                                       Pro Ala Gly Glu Met Val Thr Val Thr Gly Tr - #p Gly Arg Leu Ser Glu                            85 - #                 90 - #                 95              - - AAT ACT TCT GTT CCC TCG CCA TCA ACT CTT CA - #A GGA GTT ACA GTA CCA          336                                                                       Asn Thr Ser Val Pro Ser Pro Ser Thr Leu Gl - #n Gly Val Thr Val Pro                       100      - #           105      - #           110                  - - GTT GTA AGT AAT TCG GAA TGT CAA CAA CAA TT - #G CAA AAT CAG ACA ATC          384                                                                       Val Val Ser Asn Ser Glu Cys Gln Gln Gln Le - #u Gln Asn Gln Thr Ile                   115          - #       120          - #       125                      - - ACT GAC AAT ATG TTT TGT GCT GGT GAA TTA GA - #A GGA GGA AAG GAC TCT          432                                                                       Thr Asp Asn Met Phe Cys Ala Gly Glu Leu Gl - #u Gly Gly Lys Asp Ser               130              - #   135              - #   140                          - - T                  - #                  - #                  - #                  433                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:153:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 144 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:                             - - Val Thr Ala Ala His Cys Phe Tyr Gly Thr Le - #u Phe Pro Ile Gly Phe        1               5 - #                 10 - #                 15              - - Ser Ala Arg Ala Gly Ser Ser Thr Val Asn Se - #r Gly Gly Thr Val His                   20     - #             25     - #             30                  - - Thr Ile Leu Tyr Trp Tyr Ile His Pro Asn Ty - #r Asp Ser Gln Ser Thr               35         - #         40         - #         45                      - - Asp Phe Asp Val Ser Val Val Arg Leu Leu Se - #r Ser Leu Asn Leu Asn           50             - #     55             - #     60                          - - Gly Gly Ser Ile Arg Pro Ala Arg Leu Val As - #p Ser Gly Thr Asp Leu       65                 - # 70                 - # 75                 - # 80       - - Pro Ala Gly Glu Met Val Thr Val Thr Gly Tr - #p Gly Arg Leu Ser Glu                       85 - #                 90 - #                 95              - - Asn Thr Ser Val Pro Ser Pro Ser Thr Leu Gl - #n Gly Val Thr Val Pro                  100      - #           105      - #           110                  - - Val Val Ser Asn Ser Glu Cys Gln Gln Gln Le - #u Gln Asn Gln Thr Ile              115          - #       120          - #       125                      - - Thr Asp Asn Met Phe Cys Ala Gly Glu Leu Gl - #u Gly Gly Lys Asp Ser          130              - #   135              - #   140                          - -  - - (2) INFORMATION FOR SEQ ID NO:154:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 426 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..426                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:                             - - GCC ACG ACA CCA AAT TCG AAC CTG AAG GTG CG - #T TTG GGC GAA TGG GAC           48                                                                       Ala Thr Thr Pro Asn Ser Asn Leu Lys Val Ar - #g Leu Gly Glu Trp Asp             1               5 - #                 10 - #                 15              - - GTT CGC GAC CAC GAT GAG CGA CTG AAC CAC GA - #G GAA TAC GCA ATC GAA           96                                                                       Val Arg Asp His Asp Glu Arg Leu Asn His Gl - #u Glu Tyr Ala Ile Glu                        20     - #             25     - #             30                  - - CGC AAA GAA GTT CAT CCT TCA TAT TCA CCA AC - #C GAT TTC CGG AAT GAT          144                                                                       Arg Lys Glu Val His Pro Ser Tyr Ser Pro Th - #r Asp Phe Arg Asn Asp                    35         - #         40         - #         45                      - - GTA GCC TTA GTG AAA CTC GAT AGA ACT GTT AT - #T TTC AAA CAA CAT ATT          192                                                                       Val Ala Leu Val Lys Leu Asp Arg Thr Val Il - #e Phe Lys Gln His Ile                50             - #     55             - #     60                          - - TTA CCT GTC TGC TTA CCT CAT AAG CAA ATG AA - #A CTG GCT GGA AAA ATG          240                                                                       Leu Pro Val Cys Leu Pro His Lys Gln Met Ly - #s Leu Ala Gly Lys Met            65                 - # 70                 - # 75                 - # 80       - - GCA ACA GTC GCC GGA TGG GGA CGG ACG AGG CA - #C GGG CAG AGC ACT GTG          288                                                                       Ala Thr Val Ala Gly Trp Gly Arg Thr Arg Hi - #s Gly Gln Ser Thr Val                            85 - #                 90 - #                 95              - - CCG GCT GTC TTA CAA GAA GTC GAT GTC GAG GT - #G ATT CCG AAT GAA AGA          336                                                                       Pro Ala Val Leu Gln Glu Val Asp Val Glu Va - #l Ile Pro Asn Glu Arg                       100      - #           105      - #           110                  - - TGC CAG AGG TGG TTC CGT GCT GCG GGT CGA CG - #A GAA ACC ATT CAC GAT          384                                                                       Cys Gln Arg Trp Phe Arg Ala Ala Gly Arg Ar - #g Glu Thr Ile His Asp                   115          - #       120          - #       125                      - - GTC TTT CTC TGC GCC GGA TAT AAA GAG GGT GG - #T CGT GAT TCA                 - # 426                                                                    Val Phe Leu Cys Ala Gly Tyr Lys Glu Gly Gl - #y Arg Asp Ser                       130              - #   135              - #   140                          - -  - - (2) INFORMATION FOR SEQ ID NO:155:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 142 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:                             - - Ala Thr Thr Pro Asn Ser Asn Leu Lys Val Ar - #g Leu Gly Glu Trp Asp        1               5 - #                 10 - #                 15              - - Val Arg Asp His Asp Glu Arg Leu Asn His Gl - #u Glu Tyr Ala Ile Glu                   20     - #             25     - #             30                  - - Arg Lys Glu Val His Pro Ser Tyr Ser Pro Th - #r Asp Phe Arg Asn Asp               35         - #         40         - #         45                      - - Val Ala Leu Val Lys Leu Asp Arg Thr Val Il - #e Phe Lys Gln His Ile           50             - #     55             - #     60                          - - Leu Pro Val Cys Leu Pro His Lys Gln Met Ly - #s Leu Ala Gly Lys Met       65                 - # 70                 - # 75                 - # 80       - - Ala Thr Val Ala Gly Trp Gly Arg Thr Arg Hi - #s Gly Gln Ser Thr Val                       85 - #                 90 - #                 95              - - Pro Ala Val Leu Gln Glu Val Asp Val Glu Va - #l Ile Pro Asn Glu Arg                  100      - #           105      - #           110                  - - Cys Gln Arg Trp Phe Arg Ala Ala Gly Arg Ar - #g Glu Thr Ile His Asp              115          - #       120          - #       125                      - - Val Phe Leu Cys Ala Gly Tyr Lys Glu Gly Gl - #y Arg Asp Ser                  130              - #   135              - #   140                          - -  - - (2) INFORMATION FOR SEQ ID NO:156:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 390 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..390                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:                             - - GTA TAT GGC AGA GCA ACT CCA TCA CTG TTC AC - #C GTT GTA TCA GGC GCA           48                                                                       Val Tyr Gly Arg Ala Thr Pro Ser Leu Phe Th - #r Val Val Ser Gly Ala             1               5 - #                 10 - #                 15              - - CTT TAC TTA ACT GAA GGT GGA GAA CAT CAT GC - #T GTT GCT TCC ATC AAA           96                                                                       Leu Tyr Leu Thr Glu Gly Gly Glu His His Al - #a Val Ala Ser Ile Lys                        20     - #             25     - #             30                  - - TAT CAC GAA AAG TAC AGC CCA AAC ACT TTG GA - #C AAT GAT GTG GCA GTT          144                                                                       Tyr His Glu Lys Tyr Ser Pro Asn Thr Leu As - #p Asn Asp Val Ala Val                    35         - #         40         - #         45                      - - TTG AAG TTG AAA CAG CCA TTG ACT TTC AAT GC - #T AAC CAG AAA CCT GTC          192                                                                       Leu Lys Leu Lys Gln Pro Leu Thr Phe Asn Al - #a Asn Gln Lys Pro Val                50             - #     55             - #     60                          - - GCC TTG GCC TCA AAG GAT ACA CCT GGA GAC CT - #C AAA TGC AAA TTC TCT          240                                                                       Ala Leu Ala Ser Lys Asp Thr Pro Gly Asp Le - #u Lys Cys Lys Phe Ser            65                 - # 70                 - # 75                 - # 80       - - GGT TGG GGA TTA GAC GCA TAT CCA AGT GAT GT - #T TTA CCA AAT CAC TTA          288                                                                       Gly Trp Gly Leu Asp Ala Tyr Pro Ser Asp Va - #l Leu Pro Asn His Leu                            85 - #                 90 - #                 95              - - CAA AAA ATG GAT GTT CTG ACC TAC AAT AAC GC - #T GAC TGC CAA AAG TTC          336                                                                       Gln Lys Met Asp Val Leu Thr Tyr Asn Asn Al - #a Asp Cys Gln Lys Phe                       100      - #           105      - #           110                  - - CAT AAT GCT GGA CCT AAA TCT AAC ACA ATC TA - #C CCA GGA ATG TTG TGC          384                                                                       His Asn Ala Gly Pro Lys Ser Asn Thr Ile Ty - #r Pro Gly Met Leu Cys                   115          - #       120          - #       125                      - - GGA TTC                - #                  - #                  -      #          390                                                                  Gly Phe                                                                           130                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:157:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 130 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:                             - - Val Tyr Gly Arg Ala Thr Pro Ser Leu Phe Th - #r Val Val Ser Gly        Ala                                                                               1               5 - #                 10 - #                 15             - - Leu Tyr Leu Thr Glu Gly Gly Glu His His Al - #a Val Ala Ser Ile Lys                   20     - #             25     - #             30                  - - Tyr His Glu Lys Tyr Ser Pro Asn Thr Leu As - #p Asn Asp Val Ala Val               35         - #         40         - #         45                      - - Leu Lys Leu Lys Gln Pro Leu Thr Phe Asn Al - #a Asn Gln Lys Pro Val           50             - #     55             - #     60                          - - Ala Leu Ala Ser Lys Asp Thr Pro Gly Asp Le - #u Lys Cys Lys Phe Ser       65                 - # 70                 - # 75                 - # 80       - - Gly Trp Gly Leu Asp Ala Tyr Pro Ser Asp Va - #l Leu Pro Asn His Leu                       85 - #                 90 - #                 95              - - Gln Lys Met Asp Val Leu Thr Tyr Asn Asn Al - #a Asp Cys Gln Lys Phe                  100      - #           105      - #           110                  - - His Asn Ala Gly Pro Lys Ser Asn Thr Ile Ty - #r Pro Gly Met Leu Cys              115          - #       120          - #       125                      - - Gly Phe                                                                      130                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:158:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 197 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..197                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:                             - - GT GGT GGG CTG CCT TCT TCA ATA ACA TTA GCG - # CGA GTC AGA CTC GGC            47                                                                          Gly Gly Leu Pro Ser Ser Ile Thr Leu - #Ala Arg Val Arg Leu Gly                  1             - #  5                - #  10                - #  15        - - GAA CAT AAT GAT CAA TCG GGT ATA GAT TGC GA - #G GAC GAT GTT TGC GCA           95                                                                       Glu His Asn Asp Gln Ser Gly Ile Asp Cys Gl - #u Asp Asp Val Cys Ala                            20 - #                 25 - #                 30              - - GAA CCT GTC CAA GAT TTC GAT CCT GTG AAA AT - #A ATT CCA CAT CCA GAA          143                                                                       Glu Pro Val Gln Asp Phe Asp Pro Val Lys Il - #e Ile Pro His Pro Glu                        35     - #             40     - #             45                  - - TAC AAA GAC GAA CTA TTT AAA CAT GAT ATA GC - #T CTG ATA AAA TTG GTA          191                                                                       Tyr Lys Asp Glu Leu Phe Lys His Asp Ile Al - #a Leu Ile Lys Leu Val                    50         - #         55         - #         60                      - - GAA AAT                - #                  - #                  -      #          197                                                                  Glu Asn                                                                            65                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:159:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 65 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159:                             - - Gly Gly Leu Pro Ser Ser Ile Thr Leu Ala Ar - #g Val Arg Leu Gly        Glu                                                                               1               5 - #                 10 - #                 15             - - His Asn Asp Gln Ser Gly Ile Asp Cys Glu As - #p Asp Val Cys Ala Glu                   20     - #             25     - #             30                  - - Pro Val Gln Asp Phe Asp Pro Val Lys Ile Il - #e Pro His Pro Glu Tyr               35         - #         40         - #         45                      - - Lys Asp Glu Leu Phe Lys His Asp Ile Ala Le - #u Ile Lys Leu Val Glu           50             - #     55             - #     60                          - - Asn                                                                       65                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:160:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 261 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..261                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:                             - - CCG ACT ACC TTA GGC GGA TCT ACT GGT CAC AG - #C AAT GAT ATT GCC CTA           48                                                                       Pro Thr Thr Leu Gly Gly Ser Thr Gly His Se - #r Asn Asp Ile Ala Leu             1               5 - #                 10 - #                 15              - - ATC AAA GTC GAT AGA GAT ATC AAA TTC AGC AA - #A ACT GTC CAA CCT ATC           96                                                                       Ile Lys Val Asp Arg Asp Ile Lys Phe Ser Ly - #s Thr Val Gln Pro Ile                        20     - #             25     - #             30                  - - AAA TTG CAC AAA AGT TTA ATA AAT GGA GGT GA - #A AAA TTG AAA ATT ACT          144                                                                       Lys Leu His Lys Ser Leu Ile Asn Gly Gly Gl - #u Lys Leu Lys Ile Thr                    35         - #         40         - #         45                      - - GGA TGG GGA TTG ACG AAT CAA AGT CAT AGT GA - #T GAA CCA GAT GTT CTT          192                                                                       Gly Trp Gly Leu Thr Asn Gln Ser His Ser As - #p Glu Pro Asp Val Leu                50             - #     55             - #     60                          - - CAA GAG TTG CAT GTA AAA GCA CTT ACT GAT TC - #T GAG TGC GAG AAA GCT          240                                                                       Gln Glu Leu His Val Lys Ala Leu Thr Asp Se - #r Glu Cys Glu Lys Ala            65                 - # 70                 - # 75                 - # 80       - - ACA GGT GAA GAC CAT CCT ACA       - #                  - #                     261                                                                     Thr Gly Glu Asp His Pro Thr                                                                    85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:161:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 87 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:                             - - Pro Thr Thr Leu Gly Gly Ser Thr Gly His Se - #r Asn Asp Ile Ala Leu        1               5 - #                 10 - #                 15              - - Ile Lys Val Asp Arg Asp Ile Lys Phe Ser Ly - #s Thr Val Gln Pro Ile                   20     - #             25     - #             30                  - - Lys Leu His Lys Ser Leu Ile Asn Gly Gly Gl - #u Lys Leu Lys Ile Thr               35         - #         40         - #         45                      - - Gly Trp Gly Leu Thr Asn Gln Ser His Ser As - #p Glu Pro Asp Val Leu           50             - #     55             - #     60                          - - Gln Glu Leu His Val Lys Ala Leu Thr Asp Se - #r Glu Cys Glu Lys Ala       65                 - # 70                 - # 75                 - # 80       - - Thr Gly Glu Asp His Pro Thr                                                               85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:162:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 341 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..341                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:                             - - GA AAA TCT CGA ATT GGG CGA CAG CTT AAA ATT - # CAC TAT GGA AAT AAC            47                                                                          Lys Ser Arg Ile Gly Arg Gln Leu Lys - #Ile His Tyr Gly Asn Asn                  1             - #  5                - #  10                - #  15        - - GAC TGG CAC TTT GGC TTC GTT AGT ATT GTA AA - #A AAG GCT ATT ATT CAT           95                                                                       Asp Trp His Phe Gly Phe Val Ser Ile Val Ly - #s Lys Ala Ile Ile His                            20 - #                 25 - #                 30              - - CCA AAT TAC AAC CCA GTG ACA TTT GAT AGT GA - #T GTG GCC CTT CTG AAG          143                                                                       Pro Asn Tyr Asn Pro Val Thr Phe Asp Ser As - #p Val Ala Leu Leu Lys                        35     - #             40     - #             45                  - - CTG CAC TCT CCA ATT ACC TTC ACA AAT GGC GT - #T CAT AAA GTG TCG CTG          191                                                                       Leu His Ser Pro Ile Thr Phe Thr Asn Gly Va - #l His Lys Val Ser Leu                    50         - #         55         - #         60                      - - GTC GAA AAA GGT CAA GAT CCT GTA CCT TAT TC - #A CCT GCG ATG ATC ACT          239                                                                       Val Glu Lys Gly Gln Asp Pro Val Pro Tyr Se - #r Pro Ala Met Ile Thr                65             - #     70             - #     75                          - - GGC TGG GGC CAT ACA ATG GAA GGT GAT ACT AG - #T ATT TCG CAA ATT TTA          287                                                                       Gly Trp Gly His Thr Met Glu Gly Asp Thr Se - #r Ile Ser Gln Ile Leu            80                 - # 85                 - # 90                 - # 95       - - CAA GGA GCT GTG GTC CCA ATC GTA AAC AGA AA - #T GAT TGT CCG AAT TAT          335                                                                       Gln Gly Ala Val Val Pro Ile Val Asn Arg As - #n Asp Cys Pro Asn Tyr                           100  - #               105  - #               110              - - GGA CTC                - #                  - #                  -      #          341                                                                  Gly Leu                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:163:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 113 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163:                             - - Lys Ser Arg Ile Gly Arg Gln Leu Lys Ile Hi - #s Tyr Gly Asn Asn        Asp                                                                               1               5 - #                 10 - #                 15             - - Trp His Phe Gly Phe Val Ser Ile Val Lys Ly - #s Ala Ile Ile His Pro                   20     - #             25     - #             30                  - - Asn Tyr Asn Pro Val Thr Phe Asp Ser Asp Va - #l Ala Leu Leu Lys Leu               35         - #         40         - #         45                      - - His Ser Pro Ile Thr Phe Thr Asn Gly Val Hi - #s Lys Val Ser Leu Val           50             - #     55             - #     60                          - - Glu Lys Gly Gln Asp Pro Val Pro Tyr Ser Pr - #o Ala Met Ile Thr Gly       65                 - # 70                 - # 75                 - # 80       - - Trp Gly His Thr Met Glu Gly Asp Thr Ser Il - #e Ser Gln Ile Leu Gln                       85 - #                 90 - #                 95              - - Gly Ala Val Val Pro Ile Val Asn Arg Asn As - #p Cys Pro Asn Tyr Gly                  100      - #           105      - #           110                  - - Leu                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:164:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 267 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..267                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164:                             - - CCG ACT ACC TTA GGC GGA TCT ACT GGT CAC AG - #C AAT GAT ATT GCC CTA           48                                                                       Pro Thr Thr Leu Gly Gly Ser Thr Gly His Se - #r Asn Asp Ile Ala Leu             1               5 - #                 10 - #                 15              - - ATC AAA GTC GAT AGA GAT ATC AAA TTC AGC AA - #A ACT GTC CAA CCT ATC           96                                                                       Ile Lys Val Asp Arg Asp Ile Lys Phe Ser Ly - #s Thr Val Gln Pro Ile                        20     - #             25     - #             30                  - - AAA TTG CAC AAA AGT TTA ATA AAT GGA GGT GA - #A AAA TTG AAA ATT ACT          144                                                                       Lys Leu His Lys Ser Leu Ile Asn Gly Gly Gl - #u Lys Leu Lys Ile Thr                    35         - #         40         - #         45                      - - GGA TGG GGA TTG ACG AAT CAA AGT CAT AGT GA - #T GAA CCA GAT GTT CTT          192                                                                       Gly Trp Gly Leu Thr Asn Gln Ser His Ser As - #p Glu Pro Asp Val Leu                50             - #     55             - #     60                          - - CAA GAG TTG CAT GTA AAA GCA CTT ACT GAT TC - #T GAG TGC GAG AAA GCT          240                                                                       Gln Glu Leu His Val Lys Ala Leu Thr Asp Se - #r Glu Cys Glu Lys Ala            65                 - # 70                 - # 75                 - # 80       - - ACA GGT GAA GAC CAT CCT ACA CAC CTT    - #                  - #                267                                                                     Thr Gly Glu Asp His Pro Thr His Leu                                                            85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:165:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 89 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:                             - - Pro Thr Thr Leu Gly Gly Ser Thr Gly His Se - #r Asn Asp Ile Ala Leu        1               5 - #                 10 - #                 15              - - Ile Lys Val Asp Arg Asp Ile Lys Phe Ser Ly - #s Thr Val Gln Pro Ile                   20     - #             25     - #             30                  - - Lys Leu His Lys Ser Leu Ile Asn Gly Gly Gl - #u Lys Leu Lys Ile Thr               35         - #         40         - #         45                      - - Gly Trp Gly Leu Thr Asn Gln Ser His Ser As - #p Glu Pro Asp Val Leu           50             - #     55             - #     60                          - - Gln Glu Leu His Val Lys Ala Leu Thr Asp Se - #r Glu Cys Glu Lys Ala       65                 - # 70                 - # 75                 - # 80       - - Thr Gly Glu Asp His Pro Thr His Leu                                                       85                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:166:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166:                             - - CCCAAATTTT CCATWGCNCC NGC           - #                  - #                    23                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:167:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:                             - - Leu Ala Thr Thr Gln Phe Gln Ala Thr His Al - #a Arg Ser Ala Phe Pro      1               5   - #                10  - #                15               - - Cys Phe Asp Glu Pro Ala Met                                                          20                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:168:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1..20                                                           (D) OTHER INFORMATION: - #/label= primer                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:                             - - CAATTYCAAG CTACYCATGC            - #                  - #                      - # 20                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:169:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 383 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..382                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:                             - - C CGT AGT GCT TTC CCT TGT TTC GAT GAA CCA - #GCA ATG AAG GCC CAT             46                                                                          Arg Ser Ala Phe Pro Cys Phe Asp Glu P - #ro Ala Met Lys Ala His                 1              - # 5                 - # 10                 - # 15         - - TTC GAA ATC AGC CTT ATA CAC CAT GAA AAA TT - #G AAA GCA ATT TCC AAT           94                                                                       Phe Glu Ile Ser Leu Ile His His Glu Lys Le - #u Lys Ala Ile Ser Asn                            20 - #                 25 - #                 30              - - ATG GGT GTA GCA AAG GAA GAA AAC TTA GAT AA - #C AAC CGA AAA AGA ACA          142                                                                       Met Gly Val Ala Lys Glu Glu Asn Leu Asp As - #n Asn Arg Lys Arg Thr                        35     - #             40     - #             45                  - - ACA TTC GAA CAA TCA GTT CTC ATG TCT CCA TA - #C CTG GTG GCG TTT ATT          190                                                                       Thr Phe Glu Gln Ser Val Leu Met Ser Pro Ty - #r Leu Val Ala Phe Ile                    50         - #         55         - #         60                      - - ATC TCA GAT TTC GAA TAT GTA GAA AAA ATT TC - #A GGA CCA GTG AAA TAC          238                                                                       Ile Ser Asp Phe Glu Tyr Val Glu Lys Ile Se - #r Gly Pro Val Lys Tyr                65             - #     70             - #     75                          - - AGA ATA TAT ACT GAT CCT TTC TCG ATT GAT CA - #A GCT GAC TAT GCA TTG          286                                                                       Arg Ile Tyr Thr Asp Pro Phe Ser Ile Asp Gl - #n Ala Asp Tyr Ala Leu            80                 - # 85                 - # 90                 - # 95       - - ACT ATG AGC CCC AAA AAT TTT AAC GGC TTT GG - #A ACA ACT CAC AGG TGT          334                                                                       Thr Met Ser Pro Lys Asn Phe Asn Gly Phe Gl - #y Thr Thr His Arg Cys                           100  - #               105  - #               110              - - AAA ATA TGT TTT GAA CAA GTT GGA CCA AGC AG - #C CAT TCC AGA TTT TGC          382                                                                       Lys Ile Cys Phe Glu Gln Val Gly Pro Ser Se - #r His Ser Arg Phe Cys                       115      - #           120      - #           125                  - - T                  - #                  - #                  - #                  383                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:170:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 127 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170:                             - - Arg Ser Ala Phe Pro Cys Phe Asp Glu Pro Al - #a Met Lys Ala His Phe        1               5 - #                 10 - #                 15              - - Glu Ile Ser Leu Ile His His Glu Lys Leu Ly - #s Ala Ile Ser Asn Met                   20     - #             25     - #             30                  - - Gly Val Ala Lys Glu Glu Asn Leu Asp Asn As - #n Arg Lys Arg Thr Thr               35         - #         40         - #         45                      - - Phe Glu Gln Ser Val Leu Met Ser Pro Tyr Le - #u Val Ala Phe Ile Ile           50             - #     55             - #     60                          - - Ser Asp Phe Glu Tyr Val Glu Lys Ile Ser Gl - #y Pro Val Lys Tyr Arg       65                 - # 70                 - # 75                 - # 80       - - Ile Tyr Thr Asp Pro Phe Ser Ile Asp Gln Al - #a Asp Tyr Ala Leu Thr                       85 - #                 90 - #                 95              - - Met Ser Pro Lys Asn Phe Asn Gly Phe Gly Th - #r Thr His Arg Cys Lys                  100      - #           105      - #           110                  - - Ile Cys Phe Glu Gln Val Gly Pro Ser Ser Hi - #s Ser Arg Phe Cys                  115          - #       120          - #       125                      - -  - - (2) INFORMATION FOR SEQ ID NO:171:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 537 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..535                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:                             - - A TAT ATT TTG GAA ATA AAA GGC TTC AAA TCT - #ACT TTG AGA GAT GAC             46                                                                          Tyr Ile Leu Glu Ile Lys Gly Phe Lys S - #er Thr Leu Arg Asp Asp                 1              - # 5                 - # 10                 - # 15         - - ATG GCT GGC TTC TAC AAA AGT TCA TAC AAA AC - #G CCA AAA GGA GAA ACA           94                                                                       Met Ala Gly Phe Tyr Lys Ser Ser Tyr Lys Th - #r Pro Lys Gly Glu Thr                            20 - #                 25 - #                 30              - - AGA TGG TTG GCT ACA ACC CAG TTT CAG GCA AC - #T CAT GCC CGT AGT GCT          142                                                                       Arg Trp Leu Ala Thr Thr Gln Phe Gln Ala Th - #r His Ala Arg Ser Ala                        35     - #             40     - #             45                  - - TTC CCT TGT TTC GAT GAA CCA GCA ATG AAG GC - #C CAT TTC GAA ATC AGC          190                                                                       Phe Pro Cys Phe Asp Glu Pro Ala Met Lys Al - #a His Phe Glu Ile Ser                    50         - #         55         - #         60                      - - CTT ATA CAC CAT GAA AAA TTG AAA GCA ATT TC - #C AAT ATG GGT GTA GCA          238                                                                       Leu Ile His His Glu Lys Leu Lys Ala Ile Se - #r Asn Met Gly Val Ala                65             - #     70             - #     75                          - - AAG GAA GAA AAC TTA GAT AAC AAC CGA AAA AG - #A ACA ACA TTC GAA CAA          286                                                                       Lys Glu Glu Asn Leu Asp Asn Asn Arg Lys Ar - #g Thr Thr Phe Glu Gln            80                 - # 85                 - # 90                 - # 95       - - TCA GTT CTC ATG TCT CCA TAC CTG GTG GCG TT - #T ATT ATC TCA GAT TTC          334                                                                       Ser Val Leu Met Ser Pro Tyr Leu Val Ala Ph - #e Ile Ile Ser Asp Phe                           100  - #               105  - #               110              - - GAA TAT GTA GAA AAA ATT TCA GGA CCA GTG AA - #A TAC AGA ATA TAT ACT          382                                                                       Glu Tyr Val Glu Lys Ile Ser Gly Pro Val Ly - #s Tyr Arg Ile Tyr Thr                       115      - #           120      - #           125                  - - GAT CCT TTC TCG ATT GAT CAA GCT GAC TAT GC - #A TTG ACT ATG AGC CCC          430                                                                       Asp Pro Phe Ser Ile Asp Gln Ala Asp Tyr Al - #a Leu Thr Met Ser Pro                   130          - #       135          - #       140                      - - AAA AAT TTT AAC GGC TTT GGA ACA ACT CAC AG - #G TGT AAA ATA TGT TTT          478                                                                       Lys Asn Phe Asn Gly Phe Gly Thr Thr His Ar - #g Cys Lys Ile Cys Phe               145              - #   150              - #   155                          - - GAA CAA GTT GGA CCA AGC AGC CAT TCC AGA TT - #T TGC TGC CGG CGC AAT          526                                                                       Glu Gln Val Gly Pro Ser Ser His Ser Arg Ph - #e Cys Cys Arg Arg Asn           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - GGA AAA TTT GG             - #                  - #                      - #      537                                                                 Gly Lys Phe                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:172:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 178 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:172:                             - - Tyr Ile Leu Glu Ile Lys Gly Phe Lys Ser Th - #r Leu Arg Asp Asp Met        1               5 - #                 10 - #                 15              - - Ala Gly Phe Tyr Lys Ser Ser Tyr Lys Thr Pr - #o Lys Gly Glu Thr Arg                   20     - #             25     - #             30                  - - Trp Leu Ala Thr Thr Gln Phe Gln Ala Thr Hi - #s Ala Arg Ser Ala Phe               35         - #         40         - #         45                      - - Pro Cys Phe Asp Glu Pro Ala Met Lys Ala Hi - #s Phe Glu Ile Ser Leu           50             - #     55             - #     60                          - - Ile His His Glu Lys Leu Lys Ala Ile Ser As - #n Met Gly Val Ala Lys       65                 - # 70                 - # 75                 - # 80       - - Glu Glu Asn Leu Asp Asn Asn Arg Lys Arg Th - #r Thr Phe Glu Gln Ser                       85 - #                 90 - #                 95              - - Val Leu Met Ser Pro Tyr Leu Val Ala Phe Il - #e Ile Ser Asp Phe Glu                  100      - #           105      - #           110                  - - Tyr Val Glu Lys Ile Ser Gly Pro Val Lys Ty - #r Arg Ile Tyr Thr Asp              115          - #       120          - #       125                      - - Pro Phe Ser Ile Asp Gln Ala Asp Tyr Ala Le - #u Thr Met Ser Pro Lys          130              - #   135              - #   140                          - - Asn Phe Asn Gly Phe Gly Thr Thr His Arg Cy - #s Lys Ile Cys Phe Glu      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gln Val Gly Pro Ser Ser His Ser Arg Phe Cy - #s Cys Arg Arg Asn        Gly                                                                                             165  - #               170  - #               175             - - Lys Phe                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:173:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 25 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173:                             - - TTGGGATACA CTTTGACTGT TAACC          - #                  - #                   25                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:174:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174:                             - - GTGAGCAACC ATTATTTCCA TATC          - #                  - #                    24                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:175:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1..23                                                           (D) OTHER INFORMATION: - #/label= primer                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175:                             - - CTTGTACGAT TGTCTCAACA GGC           - #                  - #                    23                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:176:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1..27                                                           (D) OTHER INFORMATION: - #/label= primer                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:176:                             - - CTTTCCTCAC AATACCACCA AGGAAGC          - #                  - #                 27                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:177:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 573 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..573                                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:177:                             - - CGA TCT TTT AGA CAA GCG AAA TTG ATA ACG AA - #G TTT TCG AAG TCG GAT           48                                                                       Arg Ser Phe Arg Gln Ala Lys Leu Ile Thr Ly - #s Phe Ser Lys Ser Asp             1               5 - #                 10 - #                 15              - - GAA GTA AAA ACC TTG CGT TGG TTT CCC CGG TC - #C CAG GAT CAG GAA CAG           96                                                                       Glu Val Lys Thr Leu Arg Trp Phe Pro Arg Se - #r Gln Asp Gln Glu Gln                        20     - #             25     - #             30                  - - TTG CAC TTT ACC CCA ATG AGG GAA TTC GTG CA - #T CCC CAT TTT ACC GAA          144                                                                       Leu His Phe Thr Pro Met Arg Glu Phe Val Hi - #s Pro His Phe Thr Glu                    35         - #         40         - #         45                      - - CAT ATT GAT GAA GAA TTC CAC CGA TTC ATC AA - #T AAA CAC GGA AAA ATT          192                                                                       His Ile Asp Glu Glu Phe His Arg Phe Ile As - #n Lys His Gly Lys Ile                50             - #     55             - #     60                          - - TAT AAT AAA AAT GAA GAA CAT CAT TTC CGC AA - #A GAA ATT TTC AGA CTA          240                                                                       Tyr Asn Lys Asn Glu Glu His His Phe Arg Ly - #s Glu Ile Phe Arg Leu            65                 - # 70                 - # 75                 - # 80       - - AAC TTG AGG TAC ATT TTT TCT AAG AAT CGT GC - #A AAT TTG GGA TAC ACT          288                                                                       Asn Leu Arg Tyr Ile Phe Ser Lys Asn Arg Al - #a Asn Leu Gly Tyr Thr                            85 - #                 90 - #                 95              - - TTG ACT GTT AAC CAT TTG GCT GAT CGT ACT GA - #A GCT GAA CTT AAG GCT          336                                                                       Leu Thr Val Asn His Leu Ala Asp Arg Thr Gl - #u Ala Glu Leu Lys Ala                       100      - #           105      - #           110                  - - TTG AGA GGA CAC AGA CCT TCC TCC GGT TAT AA - #T GGC GGT TTA CCC TTT          384                                                                       Leu Arg Gly His Arg Pro Ser Ser Gly Tyr As - #n Gly Gly Leu Pro Phe                   115          - #       120          - #       125                      - - CCT CAC AAT ACC ACC AAG GAA GCA AGA AAT TT - #A CCA GAT TCT TTC GAC          432                                                                       Pro His Asn Thr Thr Lys Glu Ala Arg Asn Le - #u Pro Asp Ser Phe Asp               130              - #   135              - #   140                          - - TGG CGA ATT TAT GGA GCT GTT ACT CCA GTT AA - #A GAT CAA TCT GTT TGT          480                                                                       Trp Arg Ile Tyr Gly Ala Val Thr Pro Val Ly - #s Asp Gln Ser Val Cys           145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - GGT TCC TGC TGG TCT TTC GGA ACA ATT GGA GC - #A ATC GAA GGT GCA        TAT      528                                                                    Gly Ser Cys Trp Ser Phe Gly Thr Ile Gly Al - #a Ile Glu Gly Ala Tyr                          165  - #               170  - #               175              - - TTC TTG AAA ACG GCG GTA ATC TGT ACG ATG TC - #T CAC AGC TTG ATG              57 - #3                                                                   Phe Leu Lys Thr Ala Val Ile Cys Thr Met Se - #r His Ser Leu Met                           180      - #           185      - #           190                  - -  - - (2) INFORMATION FOR SEQ ID NO:178:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 191 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:178:                             - - Arg Ser Phe Arg Gln Ala Lys Leu Ile Thr Ly - #s Phe Ser Lys Ser Asp        1               5 - #                 10 - #                 15              - - Glu Val Lys Thr Leu Arg Trp Phe Pro Arg Se - #r Gln Asp Gln Glu Gln                   20     - #             25     - #             30                  - - Leu His Phe Thr Pro Met Arg Glu Phe Val Hi - #s Pro His Phe Thr Glu               35         - #         40         - #         45                      - - His Ile Asp Glu Glu Phe His Arg Phe Ile As - #n Lys His Gly Lys Ile           50             - #     55             - #     60                          - - Tyr Asn Lys Asn Glu Glu His His Phe Arg Ly - #s Glu Ile Phe Arg Leu       65                 - # 70                 - # 75                 - # 80       - - Asn Leu Arg Tyr Ile Phe Ser Lys Asn Arg Al - #a Asn Leu Gly Tyr Thr                       85 - #                 90 - #                 95              - - Leu Thr Val Asn His Leu Ala Asp Arg Thr Gl - #u Ala Glu Leu Lys Ala                  100      - #           105      - #           110                  - - Leu Arg Gly His Arg Pro Ser Ser Gly Tyr As - #n Gly Gly Leu Pro Phe              115          - #       120          - #       125                      - - Pro His Asn Thr Thr Lys Glu Ala Arg Asn Le - #u Pro Asp Ser Phe Asp          130              - #   135              - #   140                          - - Trp Arg Ile Tyr Gly Ala Val Thr Pro Val Ly - #s Asp Gln Ser Val Cys      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Gly Ser Cys Trp Ser Phe Gly Thr Ile Gly Al - #a Ile Glu Gly Ala        Tyr                                                                                             165  - #               170  - #               175             - - Phe Leu Lys Thr Ala Val Ile Cys Thr Met Se - #r His Ser Leu Met                      180      - #           185      - #           190                  - -  - - (2) INFORMATION FOR SEQ ID NO:179:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:179:                             - - Ile Val Gly Gly Glu Asp Val Asp Ile Ser Th - #r Cys Gly Trp Cys          1               5   - #                10  - #                15               - -  - - (2) INFORMATION FOR SEQ ID NO:180:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180:                             - - Ile Val Gly Gly His Asp Thr Ser Ile Lys Gl - #n His Pro Tyr Gln Val      1               5   - #                10  - #                15               - - Ser                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:181:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181:                             - - Ile Val Gly Gly Val Ser Val Asn Ile Asn As - #p Tyr Gly Tyr Gln Leu      1               5   - #                10  - #                15               - - Ser Leu Gln Ser Asn Gly Arg                                                          20                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:182:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 32 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1..32                                                           (D) OTHER INFORMATION: - #/label= primer                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182:                             - - GGACGAATTC TTAAACACCA GACACTTCCT TG       - #                  - #              32                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:183:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 32 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1..32                                                           (D) OTHER INFORMATION: - #/label= primer                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183:                             - - GAGCTCTCGA GAATCGTAGG AGGACACGAT AC       - #                  - #              32                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:184:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 34 amino - #acids                                                 (B) TYPE: amino acid                                                          (C) STRANDEDNESS:                                                             (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184:                             - - Ile Val Gly Gly Glu Asp Val Asp Ile Ser Th - #r Cys Gly Trp Gln Ile      1               5   - #                10  - #                15               - - Ser Phe Gln Ser Glu Asn Leu His Phe Cys Gl - #y Gly Ser Ile Ile Ala                  20      - #            25      - #            30                   - - Pro Lys                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:185:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 26 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1..26                                                           (D) OTHER INFORMATION: - #/label= primer                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:185:                             - - GAAGATGTWG ATATTTCWAC ATGTGG          - #                  - #                  26                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:186:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: misc.sub.-- - #feature                                          (B) LOCATION: 1..27                                                           (D) OTHER INFORMATION: - #/label= primer                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186:                             - - GAAAATGAAA TCCACTTAAA CATTACG          - #                  - #                 27                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:187:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187:                             - - CTCTTATTGT ACGAGGGATG C           - #                  - #                      - #21                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:188:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: other nucleic acid                                         (A) DESCRIPTION: /desc - #= "primer"                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188:                             - - ATTCCTCGTG GTTCAGTCGC TC           - #                  - #                     22                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:189:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 778 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA                                              - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..778                                                          (D) OTHER INFORMATION: - #/note= "At pos. bp 13, change A to                       N.  At - # pos. aa 4, substitute Xaa."                          - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:189:                             - - T GAT ACC TCA GAN TTG CCG GTA CAA AGG CGA - #ACG GTT TCG AGT GCG             46                                                                          Asp Thr Ser Xaa Leu Pro Val Gln Arg A - #rg Thr Val Ser Ser Ala                 1              - # 5                 - # 10                 - # 15         - - GTT TGT CAA TTT TCG TGC GTC CTG GGC GGC GG - #A AAA CCT CTT GAC CTG           94                                                                       Val Cys Gln Phe Ser Cys Val Leu Gly Gly Gl - #y Lys Pro Leu Asp Leu                            20 - #                 25 - #                 30              - - TGC AGC GGC GGA ATG ATC TGG TCG TGC TGC GT - #C GAC AGG GAC ATT CGG          142                                                                       Cys Ser Gly Gly Met Ile Trp Ser Cys Cys Va - #l Asp Arg Asp Ile Arg                        35     - #             40     - #             45                  - - CCT GAG CCG CAG CAC CAG GGC GCT CTG CAG AA - #C GCA ACT TGT GGA GAA          190                                                                       Pro Glu Pro Gln His Gln Gly Ala Leu Gln As - #n Ala Thr Cys Gly Glu                    50         - #         55         - #         60                      - - TTG TAC ACG AGG TCT AAT AGA ATC GTA GGA GG - #T CAT TCA ACA GGA TTC          238                                                                       Leu Tyr Thr Arg Ser Asn Arg Ile Val Gly Gl - #y His Ser Thr Gly Phe                65             - #     70             - #     75                          - - GGG TCT CAT CCT TGG CAG GCG GCT TTG ATC AA - #A TCA GGA TTT TTG AGT          286                                                                       Gly Ser His Pro Trp Gln Ala Ala Leu Ile Ly - #s Ser Gly Phe Leu Ser            80                 - # 85                 - # 90                 - # 95       - - AAA AAA TTA TCT TGC GGT GGC GCT TTA GTT AG - #C GAT CGA TGG GTT ATA          334                                                                       Lys Lys Leu Ser Cys Gly Gly Ala Leu Val Se - #r Asp Arg Trp Val Ile                           100  - #               105  - #               110              - - ACT GCT GCA CAT TGC GTT GCC ACG ACA CCA AA - #T TCG AAC CTG AAG GTG          382                                                                       Thr Ala Ala His Cys Val Ala Thr Thr Pro As - #n Ser Asn Leu Lys Val                       115      - #           120      - #           125                  - - CGA TTG GGC GAA TGG GAC GTC CGC GAC CAC GA - #T GAG CGA CTG AAC CAC          430                                                                       Arg Leu Gly Glu Trp Asp Val Arg Asp His As - #p Glu Arg Leu Asn His                   130          - #       135          - #       140                      - - GAG GAA TAC GCA ATC GAA CGC AAA GAA GTT CA - #T CCT TCA TAT TCA CCA          478                                                                       Glu Glu Tyr Ala Ile Glu Arg Lys Glu Val Hi - #s Pro Ser Tyr Ser Pro               145              - #   150              - #   155                          - - ACC GAT TTC CGG AAT GAT GTA GCC TTA GTG AA - #A CTC GAT AGA ACT GTT          526                                                                       Thr Asp Phe Arg Asn Asp Val Ala Leu Val Ly - #s Leu Asp Arg Thr Val           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - ATT TTC AAA CAA CAT ATT TTA CCT GTC TGC TT - #A CCT CAT AAG CAA        ATG      574                                                                    Ile Phe Lys Gln His Ile Leu Pro Val Cys Le - #u Pro His Lys Gln Met                          180  - #               185  - #               190              - - AAA CTG GCT GGA AAA ATG GCA ACA GTC GCC GG - #A TGG GGA CGG ACG AGG          622                                                                       Lys Leu Ala Gly Lys Met Ala Thr Val Ala Gl - #y Trp Gly Arg Thr Arg                       195      - #           200      - #           205                  - - CAC GGG CAG AGC ACT GTG CCG GCT GTC TTA CA - #A GAA GTC GAT GTC GAG          670                                                                       His Gly Gln Ser Thr Val Pro Ala Val Leu Gl - #n Glu Val Asp Val Glu                   210          - #       215          - #       220                      - - GTG ATT CCG AAT GAA AGA TGC CAG AGG TGG TT - #C CGT GCT GCG GGT CGA          718                                                                       Val Ile Pro Asn Glu Arg Cys Gln Arg Trp Ph - #e Arg Ala Ala Gly Arg               225              - #   230              - #   235                          - - CGA GAA ACC ATT CAC GAT GTC TTT CTC TGC GC - #C GGA TAT AAA GAG GGT          766                                                                       Arg Glu Thr Ile His Asp Val Phe Leu Cys Al - #a Gly Tyr Lys Glu Gly           240                 2 - #45                 2 - #50                 2 -      #55                                                                              - - GGT CGT GAT TCA            - #                  - #                      - #      778                                                                 Gly Arg Asp Ser                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:190:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 259 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:190:                             - - Asp Thr Ser Xaa Leu Pro Val Gln Arg Arg Th - #r Val Ser Ser Ala Val        1               5 - #                 10 - #                 15              - - Cys Gln Phe Ser Cys Val Leu Gly Gly Gly Ly - #s Pro Leu Asp Leu Cys                   20     - #             25     - #             30                  - - Ser Gly Gly Met Ile Trp Ser Cys Cys Val As - #p Arg Asp Ile Arg Pro               35         - #         40         - #         45                      - - Glu Pro Gln His Gln Gly Ala Leu Gln Asn Al - #a Thr Cys Gly Glu Leu           50             - #     55             - #     60                          - - Tyr Thr Arg Ser Asn Arg Ile Val Gly Gly Hi - #s Ser Thr Gly Phe Gly       65                 - # 70                 - # 75                 - # 80       - - Ser His Pro Trp Gln Ala Ala Leu Ile Lys Se - #r Gly Phe Leu Ser Lys                       85 - #                 90 - #                 95              - - Lys Leu Ser Cys Gly Gly Ala Leu Val Ser As - #p Arg Trp Val Ile Thr                  100      - #           105      - #           110                  - - Ala Ala His Cys Val Ala Thr Thr Pro Asn Se - #r Asn Leu Lys Val Arg              115          - #       120          - #       125                      - - Leu Gly Glu Trp Asp Val Arg Asp His Asp Gl - #u Arg Leu Asn His Glu          130              - #   135              - #   140                          - - Glu Tyr Ala Ile Glu Arg Lys Glu Val His Pr - #o Ser Tyr Ser Pro Thr      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Asp Phe Arg Asn Asp Val Ala Leu Val Lys Le - #u Asp Arg Thr Val        Ile                                                                                             165  - #               170  - #               175             - - Phe Lys Gln His Ile Leu Pro Val Cys Leu Pr - #o His Lys Gln Met Lys                  180      - #           185      - #           190                  - - Leu Ala Gly Lys Met Ala Thr Val Ala Gly Tr - #p Gly Arg Thr Arg His              195          - #       200          - #       205                      - - Gly Gln Ser Thr Val Pro Ala Val Leu Gln Gl - #u Val Asp Val Glu Val          210              - #   215              - #   220                          - - Ile Pro Asn Glu Arg Cys Gln Arg Trp Phe Ar - #g Ala Ala Gly Arg Arg      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Glu Thr Ile His Asp Val Phe Leu Cys Ala Gl - #y Tyr Lys Glu Gly        Gly                                                                                             245  - #               250  - #               255             - - Arg Asp Ser                                                             __________________________________________________________________________

What is claimed is:
 1. An isolated protein selected from the groupconsisting of:(a) a protein comprising an amino acid sequence selectedfrom the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ IDNO:9, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ IDNO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:33, SEQ IDNO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ IDNO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ IDNO:47, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ IDNO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67, SEQ ID NO:69, SEQ IDNO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ IDNO:81, SEQ ID NO:83, SEQ ID NO:85, SEQ ID NO:87, SEQ ID NO:89, SEQ IDNO:91, SEQ ID NO:93, SEQ ID NO:95, SEQ ID NO:97, SEQ ID NO:99, SEQ IDNO:101, SEQ ID NO:103, SEQ ID NO:105, SEQ ID NO:107, SEQ ID NO:109, SEQID NO:111, SEQ ID NO:121, SEQ ID NO:123, SEQ ID NO:125, SEQ ID NO:127,SEQ ID NO:129, SEQ ID NO:131, SEQ ID NO:133, SEQ ID NO:135, SEQ IDNO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ ID NO:143, SEQ ID NO:145, SEQID NO:147, SEQ ID NO:149, SEQ ID NO:151, SEQ ID NO:153, SEQ ID NO:155,SEQ ID NO:157, SEQ ID NO:159, SEQ ID NO:161, SEQ ID NO:163, SEQ IDNO:165, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:184, andSEQ ID NO:190; and, (b) a flea protein comprising a homologue of aprotein of (a), wherein said homologue comprises at least a 6 contiguousamino acid portion of an amino acid sequence selected from the groupconsisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ IDNO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:17, SEQ ID NO:19, SEQ IDNO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ IDNO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ IDNO:41, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ IDNO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ IDNO:69, SEQ ID NO:71, SEQ ID NO:75, SEQ ID NO:77, SEQ ID NO:79, SEQ IDNO:81, SEQ ID NO:83, SEQ ID NO:85, SEQ ID NO:87, SEQ ID NO:89, SEQ IDNO:91, SEQ ID NO:93, SEQ ID NO:95, SEQ ID NO:97, SEQ ID NO:99, SEQ IDNO:101, SEQ ID NO:103, SEQ ID NO:105, SEQ ID NO:107, SEQ ID NO:109, SEQID NO:111, SEQ ID NO:121, SEQ ID NO:123, SEQ ID NO:125, SEQ ID NO:129,SEQ ID NO:133, SEQ ID NO:135, SEQ ID NO:137, SEQ ID NO:141, SEQ IDNO:143, SEQ ID NO:145, SEQ ID NO:149, SEQ ID NO:153, SEQ ID NO:155, SEQID NO:157, SEQ ID NO:159, SEQ ID NO:161, SEQ ID NO:163, SEQ ID NO:165,SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:184, and SEQ IDNO:190.
 2. The protein of claim 1, wherein said protein is encoded by anucleic acid molecule selected from the group consisting of nfSP1(characterized by a coding strand comprising a nucleic acid sequenceselected from the group consisting of SEQ ID NO:18, SEQ ID NO:52 and SEQID NO:80), nfSP2 (characterized by a coding strand comprising a nucleicacid sequence selected from the group consisting of SEQ ID NO:20, SEQ IDNO:54 and SEQ ID NO:82), nfSP3 (characterized by a coding strandcomprising the nucleic acid sequence of SEQ ID NO:22), nfSP4(characterized by a coding strand comprising a nucleic acid sequenceselected from the group consisting of SEQ ID NO:16, SEQ ID NO:24 and SEQID NO:56), nfSP5 (characterized by a coding strand comprising a nucleicacid sequence selected from the group consisting of SEQ ID NO:26, SEQ IDNO:58, SEQ ID NO:84, SEQ ID NO:86 and SEQ ID NO:120), nfSP6(characterized by a coding strand comprising a nucleic acid sequenceselected from the group consisting of SEQ ID NO:28, SEQ ID NO:60 and SEQID NO:88), nfSP7 (characterized by a coding strand comprising a nucleicacid sequence selected from the group consisting of SEQ ID NO:30, SEQ IDNO:62 and SEQ ID NO:90), nfSP8 (characterized by a coding strandcomprising a nucleic acid sequence selected from the group consisting ofSEQ ID NO:32, SEQ ID NO:92 and SEQ ID NO:122), nfSP9 (characterized by acoding strand comprising a nucleic acid sequence selected from the groupconsisting of SEQ ID NO:34 and SEQ ID NO:94), nfSP10 (characterized by acoding strand comprising a nucleic acid sequence selected from the groupconsisting of SEQ ID NO:36 and SEQ ID NO:96), nfSP11 (characterized by acoding strand comprising a nucleic acid sequence selected from the groupconsisting of SEQ ID NO:38, SEQ ID NO:98 and SEQ ID NO:124), nfSP12(characterized by a coding strand comprising a nucleic acid sequenceselected from the group consisting of SEQ ID NO:64, SEQ ID NO:76, SEQ IDNO:100 and SEQ ID NO:126), nfSP13 (characterized by a coding strandcomprising a nucleic acid sequence selected from the group consisting ofSEQ ID NO:66, SEQ ID NO:78 and SEQ ID NO:102), nfSP14 (characterized bya coding strand comprising the nucleic acid sequence of SEQ ID NO:68),nfSP15 (characterized by a coding strand comprising a nucleic acidsequence selected from the group consisting of SEQ ID NO:70, SEQ IDNO:104 and SEQ ID NO:128), nfSP16 (characterized by a coding strandcomprising the nucleic acid sequence of SEQ ID NO:72), nfSP17(characterized by a coding strand comprising the nucleic acid sequenceof SEQ ID NO:74), nfSP18 (characterized by a coding strand comprisingthe nucleic acid sequence of SEQ ID NO:106), nfSP19 (characterized by acoding strand comprising a nucleic acid sequence selected from the groupconsisting of SEQ ID NO:108 and SEQ ID NO:130), nfSP20 (characterized bya coding strand comprising the nucleic acid sequence of SEQ ID NO:110),nfSP21 (characterized by a coding strand comprising the nucleic acidsequence of SEQ ID NO:132), nfSP23 (characterized by a coding strandcomprising the nucleic acid sequence of SEQ ID NO:134), nfSP24(characterized by a coding strand comprising the nucleic acid sequenceof SEQ ID NO:136), nfSP25 (characterized by a coding strand comprisingthe nucleic acid sequence of SEQ ID NO:138), nfSP26 (characterized by acoding strand comprising the nucleic acid sequence of SEQ ID NO:140),nfSP27 (characterized by a coding strand comprising the nucleic acidsequence of SEQ ID NO:142), nfSP28 (characterized by a coding strandcomprising the nucleic acid sequence of SEQ ID NO:144), nfSP29(characterized by a coding strand comprising the nucleic acid sequenceof SEQ ID NO:146), nfSP30 (characterized by a coding strand comprisingthe nucleic acid sequence of SEQ ID NO:148), nfSP31 (characterized by acoding strand comprising the nucleic acid sequence of SEQ ID NO:150),nfSP32 (characterized by a coding strand comprising the nucleic acidsequence of SEQ ID NO:152), nfSP33 (characterized by a coding strandcomprising a nucleic acid sequence selected from the group consisting ofSEQ ID NO:154 and SEQ ID NO:189), nfSP34 (characterized by a codingstrand comprising the nucleic acid sequence of SEQ ID NO:156), nfSP36(characterized by a coding strand comprising the nucleic acid sequenceof SEQ ID NO:158), nfSP37 (characterized by a coding strand comprisingthe nucleic acid sequence of SEQ ID NO:160), nfSP38 (characterized by acoding strand comprising the nucleic acid sequence of SEQ ID NO:162),and nfSP39 (characterized by a coding strand comprising the nucleic acidsequence of SEQ ID NO:164).
 3. The protein of claim 1, wherein saidprotein is encoded by a nucleic acid molecule selected from the groupconsisting of nfSP1₇₇₉ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:80), nfSP2₉₄₄ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:82),nfSP3₁₇₇ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:22), nfSP4₆₇₂ (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:16), nfSP5₁₅₇(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:84), nfSP5₂₁₈ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:86), nfSP5₈₀₆ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:120),nfSP6₉₃₂ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:88), nfSP894 (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:90), nfSP8₂₉₉(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:92), nfSP9₂₆₆ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:94), nfSP10₃₇₈ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:96),nfSP11₂₅₂ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:98), nfSP11₃₀₇ (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:124), nfSP12₁₄₄(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:64), nfSP12₂₂₅ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:100), nfSP12₇₅₈ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:126),nfSP13₈₅₀ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:102), nfSP14₂₁₃ (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:68), nfSP15₂₅₂(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:104), nfSP15₈₁₅ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:128), nfSP16₁₆₈ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:72),nfSP18₅₃₄ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:106), nfSP19₃₅₉ (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:108), nfSP19₈₅₅(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:130), nfSP20₈₄₁ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:110), nfSP21₅₉₅ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:132),nfSP23₄₂₃ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:134), nfSP24₄₁₀ (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:136), nfSP25₈₆₄(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:138), nfSP26₆₁₀ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:140), nfSP27₃₈₆ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:142),nfSP28₉₂₃ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:144), nfSP29₆₁₂ (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:146), nfSP30₆₄₁(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:148), nfSP31₆₂₆ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:150), nfSP32₄₃₃ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:152),nfSP33₇₇₈ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:189), nfSP33₄₂₆ (characterized by a coding strandhaving the nucleic acid sequence of SEQ ID NO:154), nfSP34₃₉₀(characterized by a coding strand having the nucleic acid sequence ofSEQ ID NO:156), nfSP36₁₉₇ (characterized by a coding strand having thenucleic acid sequence of SEQ ID NO:158), nfSP38₃₄₁ (characterized by acoding strand having the nucleic acid sequence of SEQ ID NO:162),nfSP37₂₆₁ (characterized by a coding strand having the nucleic acidsequence of SEQ ID NO:160), and nfSP39₂₆₇ (characterized by a codingstrand having the nucleic acid sequence of SEQ ID NO:164).
 4. Theprotein of claim 1, wherein said protein is encoded by a nucleic acidmolecule comprising a nucleic acid sequence selected from the groupconsisting of SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22,SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32,SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:52, SEQ ID NO:54,SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64,SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:74,SEQ ID NO:76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82, SEQ ID NO:84,SEQ ID NO:86, SEQ ID NO:88, SEQ ID NO:90, SEQ ID NO:92, SEQ ID NO:94,SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102, SEQ ID NO:104,SEQ ID NO:106, SEQ ID NO:108, SEQ ID NO:110, SEQ ID NO:120, SEQ IDNO:122, SEQ ID NO:124, SEQ ID NO:126, SEQ ID NO:128, SEQ ID NO:130, SEQID NO:132, SEQ ID NO:134, SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140,SEQ ID NO:142, SEQ ID NO:144, SEQ ID NO:146, SEQ ID NO:148, SEQ IDNO:150, SEQ ID NO:152, SEQ ID NO:154, SEQ ID NO:156, SEQ ID NO:158, SEQID NO:160, SEQ ID NO:162, SEQ ID NO:164, and SEQ ID NO:189.
 5. Theprotein of claim 1, wherein said protein has proteolytic activity.
 6. Amethod to identify a compound capable of inhibiting flea proteaseactivity, said method comprising:(a) contacting an isolated protein, asclaimed in claim 5, with a protease substrate and a putative inhibitorycompound under conditions in which, in the absence of said compound,said protein has proteolytic activity; (b) determining if said putativeinhibitory compound inhibits said activity by detecting cleavage of theprotease substrates; wherein decreased cleavage of the proteasesubstrate indicates an inhibitory compound.
 7. The method of claim 6,wherein said isolated protein is a Ctenocephalides felis serine proteaseprotein.
 8. The protein of claim 1, wherein said protein comprises anamino acid sequence selected from the group consisting of SEQ ID NO:1,SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:17, SEQ ID NO:19, SEQ IDNO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ IDNO:31, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ IDNO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ IDNO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:53, SEQ ID NO:55, SEQ IDNO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ IDNO:67, SEQ ID NO:69, SEQ ID NO:71, SEQ ID NO:73, SEQ ID NO:75, SEQ IDNO:77, SEQ ID NO:79, SEQ ID NO:81, SEQ ID NO:83, SEQ ID NO:85, SEQ IDNO:87, SEQ ID NO:89, SEQ ID NO:91, SEQ ID NO:93, SEQ ID NO:95, SEQ IDNO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, SEQ IDNO:107, SEQ ID NO:109, SEQ ID NO:111, SEQ ID NO:121, SEQ ID NO:123, SEQID NO:125, SEQ ID NO:127, SEQ ID NO:129, SEQ ID NO:131, SEQ ID NO:133,SEQ ID NO:135, SEQ ID NO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ IDNO:143, SEQ ID NO:145, SEQ ID NO:147, SEQ ID NO:149, SEQ ID NO:151, 3SEQID NO:153, SEQ ID NO:155, SEQ ID NO:157, SEQ ID NO:159, SEQ ID NO:161,SEQ ID NO:163, SEQ ID NO:165, SEQ ID NO:179, SEQ ID NO:180, SEQ IDNO:181, SEQ ID NO:184, and SEQ ID NO:190.
 9. The protein of claim 1,wherein said protein is produced by a process comprising culturing anisolated recombinant cell transformed with a nucleic acid moleculeencoding said protein to produce said protein.